研究者詳細 - 佐藤　あやの

論文 - 佐藤　あやの

分割表示 >> ／全件表示 84 件中 1 - 84 件目

Uncovering a Latent Bioactive Interleukin-6 Glycoform. 査読国際誌

Yasuhiro Kajihara, Yanbo Liu, Yuta Maki, Ryo Okamoto, Ayano Satoh, Yasuto Todokoro, Yurie Kanemitsu, Keito Otani

Angewandte Chemie (International ed. in English) 63 ( 50 ) e202411213 2024年8月

記述言語：英語掲載種別：研究論文（学術雑誌）

A bioinspired semisynthesis of human-interleukin-6 bearing N-glycan at Asn143 (143glycosyl-IL-6) was performed by intentional glycosylation effects and protein folding chemistry for regioselective peptide-backbone activation. 143Glycosyl-IL-6 is a genetically coded cytokine, but isolation was difficult owing to a tiny amount. IL6-polypeptide (1-141-position) with an intentionally inserted cysteine at 142-position was expressed in E. coli. The expressed polypeptide was treated with a chemical folding process to make a specific helices bundle conformation through native two-disulfide bonds (43-49 and 72-82). Utilizing the successfully formed free-142-cysteine, sequential conversions using cyanation of 142-cysteine, hydrazinolysis, and thioesterification created a long polypeptide (1-141)-thioester. However, the resultant polypeptide-thioester caused considerable aggregation owing to a highly hydrophobic peptide sequence. After the reduction of two-disulfide bonds of polypeptide (1-141)-thioester, an unprecedented hydrophilic N-glycan tag was inserted at the resultant cysteine thiols. The N-glycan tags greatly stabilized polypeptide-thioester. The subsequent native chemical ligation and desulfurization successfully gave a whole 143glycosyl-IL-6 polypeptide (183-amino acids). Removal of four N-glycan tags and immediate one-pot in vitro folding protocol efficiently produced the folded 143glycosyl-IL-6. The folded 143glycosyl-IL-6 exhibited potent cell proliferation activity. The combined studies with molecular dynamics simulation, semisynthesis, and bioassays predict the bioactive conformation of latent 143glycosyl-IL-6.

DOI： 10.1002/anie.202411213

Fucosyltransferase 8 (FUT8) and core fucose expression in oxidative stress response 査読国際誌

Yuki M. Kyunai, Mika Sakamoto, Mayuko Koreishi, Yoshio Tsujino, Ayano Satoh

PLOS ONE 18 ( 2 ) e0281516 - e0281516 2023年2月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Public Library of Science (PLoS)

GlycoMaple is a new tool to predict glycan structures based on the expression levels of 950 genes encoding glycan biosynthesis-related enzymes and proteins using RNA-seq data. The antioxidant response, protecting cells from oxidative stress, has been focused on because its activation may relieve pathological conditions, such as neurodegenerative diseases. Genes involved in the antioxidant response are defined within the GO:0006979 category, including 441 human genes. Fifteen genes overlap between the glycan biosynthesis-related genes defined by GlycoMaple and the antioxidant response genes defined by GO:0006979, one of which is FUT8. 5-Hydroxy-4-phenyl-butenolide (5H4PB) extracted from Chinese aromatic vinegar induces the expression of a series of antioxidant response genes that protect cells from oxidative stress via activation of the nuclear factor erythroid 2-related factor 2–antioxidant response element pathway. Here, we show that FUT8 is upregulated in both our RNA-seq data set of 5H4PB-treated cells and publicly available RNA-seq data set of cells treated with another antioxidant, sulforaphane. Applying our RNA-seq data set to GlycoMaple led to a prediction of an increase in the core fucose of N-glycan that was confirmed by flow cytometry using a fucose-binding lectin. These results suggest that FUT8 and core fucose expression may increase upon the antioxidant response.

DOI： 10.1371/journal.pone.0281516

A Primer on Deep Learning-Based Cellular Image Classification of Changes in the Spatial Distribution of the Golgi Apparatus After Experimental Manipulation 招待査読国際誌

Daisuke Takao, Yuki M. Kyunai, Yasushi Okada, Ayano Satoh

Methods in Molecular Biology 2557 275 - 285 2022年12月

担当区分：責任著者記述言語：英語掲載種別：論文集(書籍)内論文出版者・発行元：Springer US

The visual classification of cell images according to differences in the spatial patterns of subcellular structure is an important methodology in cell and developmental biology. Experimental perturbation of cell function can induce changes in the spatial distribution of organelles and their associated markers or labels. Here, we demonstrate how to achieve accurate, unbiased, high-throughput image classification using an artificial intelligence (AI) algorithm. We show that a convolutional neural network (CNN) algorithm can classify distinct patterns of Golgi images after drug or siRNA treatments, and we review our methods from cell preparation to image acquisition and CNN analysis.

DOI： 10.1007/978-1-0716-2639-9_18

An Electron Tomographic Analysis of Giantin-Deficient Golgi Proposes a New Function of the Golgin Protein Family 招待査読国際誌

Ayano Satoh, Mitsuko Hayashi-Nishino, Kunihiko Nishino

Methods in Molecular Biology 2557 235 - 246 2022年12月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：論文集(書籍)内論文出版者・発行元：Springer US

The Golgi apparatus is an organelle that mediates modifications, sorting, and transport of proteins and lipids. Golgins are a group of proteins with coiled-coil structures that localize to the Golgi and are thought to function as tethers to facilitate the docking of vesicles, Rab GTPases, and cytoskeleton components to the Golgi stack. Giantin is the longest golgin and has been thought to function as a tether for COPI vesicles along with other golgins, such as p115 and GM130. Contrary to our expectation that the loss of the tether will result in an increase in untethered COPI vesicles in the cytoplasm, our electron microscopy observations showed that the fenestrae normally present in Golgi cisternae were reduced upon Giantin knockdown. We also found that this structural change is accompanied by altered secretion of cargo proteins and cell surface glycosylation. These results indicate that there exists a correlation between Golgi structural changes caused by the loss of Giantin and Golgi function. Here, we describe electron tomography methods for the detection of structural changes in the Golgi.

DOI： 10.1007/978-1-0716-2639-9_15

Evaluation of skin sensitization based on interleukin‑2 promoter activation in Jurkat cells 査読国際誌

Taichi Nagahata, Yoshio Tsujino, Eiji Takayama, Haruka Hikasa, Ayano Satoh

Biomedical Reports 16 ( 1 ) 3 - 3 2021年11月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Spandidos Publications

Skin sensitization is an allergic reaction caused by certain chemical substances, and is an important factor to be taken into consideration when evaluating the safety of numerous types of products. Although animal testing has long been used to evaluate skin sensitization, the recent trend to regulate such testing has led to the development and use of alternative methods. Skin sensitization reactions are summarized in the form of an adverse outcome pathway consisting of four key events (KE), including covalent binding to skin proteins (KE1), keratinocyte activation (KE2), and dendritic cell activation (KE3). Equivalent alternative methods have been developed for KE1 to KE3, but no valid alternative has yet been developed for the evaluation of KE4 and T-cell activation. Current alternative methods rely on data from KE1 to KE3 to predict the effect of chemicals on skin sensitization. The addition of KE4 data is expected to improve the accuracy and reproducibility of such predictions. The aim of this study was to establish an assay to evaluate KE4 T-cell activation to supplement data on skin sensitization related to KE4. To evaluate T-cell activation, the Jurkat T-cell line stably expressing luciferase downstream of the pro-inflammatory cytokine interleukin-2 promoter was used. After exposure to known skin sensitizing agents and control substances, luciferase activity measurements revealed that this assay was valid for evaluating skin sensitization. However, two skin sensitizers known to have immunosuppressive effects on T-cells reacted negatively in this assay. The results revealed that this assay simultaneously allows for monitoring of the skin sensitization and immuno-suppressiveness of chemical substances and supplements KE4 T-cell activation data, and may thus contribute to reducing the use of animal experiments.

DOI： 10.3892/br.2021.1486

Glycoprotein Semisynthesis by Chemical Insertion of Glycosyl Asparagine Using a Bifunctional Thioacid-Mediated Strategy 査読国際誌

Kota Nomura, Yuta Maki, Ryo Okamoto, Ayano Satoh, Yasuhiro Kajihara

Journal of the American Chemical Society 143 ( 27 ) 10157 - 10167 2021年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：American Chemical Society (ACS)

Glycosylation is a major modification of secreted and cell surface proteins, and the resultant glycans show considerable heterogeneity in their structures. To understand the biological processes arising from each glycoform, the preparation of homogeneous glycoproteins is essential for extensive biological experiments. To establish a more robust and rapid synthetic route for the synthesis of homogeneous glycoproteins, we studied several key reactions based on amino thioacids. We found that diacyl disulfide coupling (DDC) formed with glycosyl asparagine thioacid and peptide thioacid yielded glycopeptides. This efficient coupling reaction enabled us to develop a new glycoprotein synthesis method, such as the bifunctional thioacid-mediated strategy, which can couple two peptides with the N- and C-termini of glycosyl asparagine thioacid. Previous glycoprotein synthesis methods required valuable glycosyl asparagine in the early stage and subsequent multiple glycoprotein synthesis routes, whereas the developed concept can generate glycoproteins within a few steps from peptide and glycosyl asparagine thioacid. Herein, we report the characterization of the DDC of amino thioacids and the efficient ability of glycosyl asparagine thioacid to be used for robust glycoprotein semisynthesis.

DOI： 10.1021/jacs.1c02601

Synthesis and characterization of conductive flexible cellulose carbon nanohorn sheets for human tissue applications 査読国際誌

Karthik Paneer Selvam, Taichi Nagahata, Kosuke Kato, Mayuko Koreishi, Toshiyuki Nakamura, Yoshimasa Nakamura, Takeshi Nishikawa, Ayano Satoh, Yasuhiko Hayashi

Biomaterials Research 24 ( 1 ) 18 - 18 2020年10月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer Science and Business Media LLC

Abstract

Background

Conductive sheets of cellulose and carbon nanomaterials and its human skin applications are an interesting research aspect as they have potential for applications for skin compatibility. Hence it is needed to explore the effects and shed light on these applications.

Method

To fabricate wearable, portable, flexible, lightweight, inexpensive, and biocompatible composite materials, carbon nanohorns (CNHs) and hydroxyethylcellulose (HEC) were used as precursors to prepare CNH-HEC (Cnh-cel) composite sheets. Cnh-cel sheets were prepared with different loading concentrations of CNHs (10, 20 50,100 mg) in 200 mg cellulose. To fabricate the bio-compatible sheets, a pristine composite of CNHs and HEC was prepared without any pretreatment of the materials.

Results

The obtained sheets possess a conductivity of 1.83 × 10^− 10 S/m and bio-compatible with human skin. Analysis for skin-compatibility was performed for Cnh-cel sheets by h-CLAT in vitro skin sensitization tests to evaluate the activation of THP-1 cells. It was found that THP-1 cells were not activated by Cnh-cel; hence Cnh-cel is a safe biomaterial for human skin. It was also found that the composite allowed only a maximum loading of 100 mg to retain the consistent geometry of free-standing sheets of < 100 μm thickness. Since CNHs have a unique arrangement of aggregates (dahlia structure), the composite is homogeneous, as verified by transmission electron microscopy (TEM) and, scanning electron microscopy (SEM), and other functional properties investigated by Raman spectroscopy, Fourier transform infrared spectroscopy (FT-IR), conductivity measurement, tensile strength measurement, and skin sensitization.

Conclusion

It can be concluded that cellulose and CNHs sheets are conductive and compatible to human skin applications.

DOI： 10.1186/s40824-020-00194-3

その他リンク： http://link.springer.com/article/10.1186/s40824-020-00194-3/fulltext.html

High-throughput screening of bioactive compounds via new catalytic reaction in the pooled mixture. 査読国際誌

Ayano Satoh, Yuta Nishina

Bioorganic & medicinal chemistry letters 29 ( 19 ) 126539 - 126539 2019年10月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

To increase the chances of finding new candidate molecules with medicinal properties, while expending less resource and effort, the present study used pooled substrates as starting materials. A bisindole compound that showed inhibitory activity was then isolated from the mixture, and the activity was improved by optimizing the substituents on the indole skeleton.

DOI： 10.1016/j.bmcl.2019.06.061

The Golgin Protein Giantin Regulates Interconnections Between Golgi Stacks 招待査読国際誌

Ayano Satoh, Mitsuko Hayashi-Nishino, Takuto Shakuno, Junko Masuda, Mayuko Koreishi, Runa Murakami, Yoshimasa Nakamura, Toshiyuki Nakamura, Naomi Abe-Kanoh, Yasuko Honjo, Joerg Malsam, Sidney Yu, Kunihiko Nishino

Frontiers in Cell and Developmental Biology 7 160 - 160 2019年8月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers Media SA

Golgins are a family of Golgi-localized long coiled-coil proteins. The major golgin function is thought to be the tethering of vesicles, membranes, and cytoskeletal elements to the Golgi. We previously showed that knockdown of one of the longest golgins, Giantin, altered the glycosylation patterns of cell surfaces and the kinetics of cargo transport, suggesting that Giantin maintains correct glycosylation through slowing down transport within the Golgi. Giantin knockdown also altered the sizes and numbers of mini Golgi stacks generated by microtubule de-polymerization, suggesting that it maintains the independence of individual Golgi stacks. Therefore, it is presumed that Golgi stacks lose their independence following Giantin knockdown, allowing easier and possibly increased transport among stacks and abnormal glycosylation. To gain structural insights into the independence of Golgi stacks, we herein performed electron tomography and 3D modeling of Golgi stacks in Giantin knockdown cells. Compared with control cells, Giantin-knockdown cells had fewer and smaller fenestrae within each cisterna. This was supported by data showing that the diffusion rate of Golgi membrane proteins is faster in Giantin-knockdown Golgi, indicating that Giantin knockdown structurally and functionally increases connectivity among Golgi cisternae and stacks. This increased connectivity suggests that contrary to the cis-golgin tether model, Giantin instead inhibits the tether and fusion of nearby Golgi cisternae and stacks, resulting in transport difficulties between stacks that may enable the correct glycosylation of proteins and lipids passing through the Golgi.

DOI： 10.3389/fcell.2019.00160

A RasGAP, DAB2IP, regulates lipid droplet homeostasis by serving as GAP toward RAB40C 査読国際誌

Xiaomin Luo, Chunman Li, Ran Tan, Xiaohui Xu, William K.K. Wu, Ayano Satoh, Tuanlao Wang, Sidney Yu

Oncotarget 8 ( 49 ) 85415 - 85427 2017年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Impact Journals, LLC

Lipid droplet (LD) homeostasis involves activities of various RAB small GTPases. Recently, we found RAB40C was one of the RAB proteins regulating LD homeostasis. RAB40C contains a unique SOCS domain that is required for clustering of LDs. However, its precise functional role in LD homeostasis and mechanism of regulation remain largely unknown. In this study, we observed over-accumulation of LDs in cells with RAB40C deleted by Crispr-Cas9 editing. RAB40C appeared to reduce LD accumulation after long term incubation of cells with oleic acid (24 hours). Unexpectedly, we found that Ras GTPase activating protein (GAP), DAB2IP, bound to RAB40C mainly via its GAP domain and could serve as RAB40C GAP. Studies involving overexpression of DAB2IP and its GAP defective mutant and siRNA depletion of DAB2IP all confirmed that DAB2IP negatively regulated the effect of RAB40C on LD homeostasis. These results provide a novel perspective on the regulation of RAB40C and implicate various signalling pathways regulated by DAB2IP, which may play a role in LD homeostasis via RAB40C.

DOI： 10.18632/oncotarget.19960

ULK1 phosphorylates Sec23A and mediates autophagy-induced inhibition of ER-to-Golgi traffic 査読国際誌

Wenjia Gan, Caiyun Zhang, Ka Yu Siu, Ayano Satoh, Julian A. Tanner, Sidney Yu

BMC Cell Biology 18 ( 1 ) 22 - 22 2017年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer Science and Business Media LLC

BACKGROUND: Autophagy is an inducible autodigestive process that allows cells to recycle proteins and other materials for survival during stress and nutrient deprived conditions. The kinase ULK1 is required to activate this process. ULK1 phosphorylates a number of target proteins and regulates many cellular processes including the early secretory pathway. Recently, ULK1 has been demonstrated to phosphorylate Sec16 and affects the transport of serotonin transporter at the ER exit sites (ERES), but whether ULK1 may affect the transport of other cargo proteins and general secretion has not been fully addressed. RESULTS: In this study, we identified Sec23A, a component of the COPII vesicle coat, as a target of ULK1 phosphorylation. Elevated autophagy, induced by amino acid starvation, rapamycin, or overexpression of ULK1 caused aggregation of the ERES, a region of the ER dedicated for the budding of COPII vesicles. Transport of cargo proteins was also inhibited under these conditions and was retained at the ERES. ULK1 phosphorylation of Sec23A reduced the interaction between Sec23A and Sec31A. We identified serine 207, serine 312 and threonine 405 on Sec23A as ULK1 phosphorylation sites. Among these residues, serine 207, when changed to phospho-deficient and phospho-mimicking mutants, most faithfully recapitulated the above-mentioned effects of ULK1 phospho-regulation. CONCLUSION: These findings identify Sec23A as a new target of ULK1 and uncover a mechanism of coordinating intracellular protein transport and autophagy.

DOI： 10.1186/s12860-017-0138-8

An endoplasmic reticulum protein, Nogo‐B, facilitates alcoholic liver disease through regulation of kupffer cell polarization 査読国際誌

Jin‐Kyu Park, Mingjie Shao, Moon Young Kim, Soon Koo Baik, Mee Yon Cho, Teruo Utsumi, Ayano Satoh, Xinsho Ouyang, Chuhan Chung, Yasuko Iwakiri

Hepatology 65 ( 5 ) 1720 - 1734 2017年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Ovid Technologies (Wolters Kluwer Health)

Nogo‐B (Reticulon 4B) is an endoplasmic reticulum (ER) resident protein that regulates ER structure and function. Because ER stress is known to induce M2 macrophage polarization, we examined whether Nogo‐B regulates M1/M2 polarization of Kupffer cells and alters the pathogenesis of alcoholic liver disease (ALD). M1 and M2 phenotypes were assessed in relation to Nogo‐B expression and disease severity in liver specimens from ALD patients (NCT01875211). Liver specimens from wild‐type (WT) and Nogo‐B knockout (KO) mice fed a control or Lieber‐DeCarli ethanol liquid diet (5% ethanol) for 6 weeks were analyzed for liver injury and steatosis. Kupffer cells isolated from WT and Nogo‐B KO mice were assessed for M1 and M2 activation. A significant positive correlation was observed between Nogo‐B positive Kupffer cells and disease severity in ALD patients (n = 30, r = 0.66, P = 0.048). Furthermore, Nogo‐B–positive Kupffer cells were correlated with M1 activation (inducible nitric oxide synthase) (r = 0.50, P = 0.05) and negatively with markers of M2 status (CD163) (r = −0.48, P = 0.07) in these patients. WT mice exhibited significantly increased liver injury (P < 0.05) and higher hepatic triglyceride levels (P < 0.01) compared with Nogo‐B KO mice in response to chronic ethanol feeding. Nogo‐B in Kupffer cells promoted M1 polarization, whereas absence of Nogo‐B increased ER stress and M2 polarization in Kupffer cells. Conclusion: Nogo‐B is permissive of M1 polarization of Kupffer cells, thereby accentuating liver injury in ALD in humans and mice. Nogo‐B in Kupffer cells may represent a new therapeutic target for ALD. (Hepatology 2017;65:1720‐1734).

DOI： 10.1002/hep.29051

<scp>COPI</scp>–<scp>TRAPPII</scp> activates Rab18 and regulates its lipid droplet association 査読国際誌

Chunman Li, Xiaomin Luo, Shan Zhao, Gavin KY Siu, Yongheng Liang, Hsiao Chang Chan, Ayano Satoh, Sidney SB Yu

The EMBO Journal 36 ( 4 ) 441 - 457 2016年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：EMBO

Abstract

The transport protein particle (TRAPP) was initially identified as a vesicle tethering factor in yeast and as a guanine nucleotide exchange factor (GEF) for Ypt1/Rab1. In mammals, structures and functions of various TRAPP complexes are beginning to be understood. We found that mammalian TRAPPII was a GEF for both Rab18 and Rab1. Inactivation of TRAPPII‐specific subunits by various methods including siRNA depletion and CRISPR–Cas9‐mediated deletion reduced lipolysis and resulted in aberrantly large lipid droplets. Recruitment of Rab18 onto lipid droplet (LD) surface was defective in TRAPPII‐deleted cells, but the localization of Rab1 on Golgi was not affected. COPI regulates LD homeostasis. We found that the previously documented interaction between TRAPPII and COPI was also required for the recruitment of Rab18 to the LD. We hypothesize that the interaction between COPI and TRAPPII helps bring TRAPPII onto LD surface, and TRAPPII, in turn, activates Rab18 and recruits it on the LD surface to facilitate its functions in LD homeostasis.

DOI： 10.15252/embj.201694866

その他リンク： https://onlinelibrary.wiley.com/doi/full-xml/10.15252/embj.201694866

Cellular distribution of injected PLGA-nanoparticles in the liver 査読国際誌

Jin-Kyu Park, Teruo Utsumi, Young-Eun Seo, Yang Deng, Ayano Satoh, William Mark Saltzman, Yasuko Iwakiri

Nanomedicine: Nanotechnology, Biology and Medicine 12 ( 5 ) 1365 - 1374 2016年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

The cellular fate of nanoparticles in the liver is not fully understood. Because the effectiveness and safety of nanoparticles in liver therapy depends on targeting nanoparticles to the right cell populations, this study aimed to determine a relative distribution of PLGA-nanoparticles (sizes 271±1.4 nm) among liver cells in vivo. We found that Kupffer cells were the major cells that took up nanoparticles, followed by liver sinusoidal endothelial cells and hepatic stellate cells. Nanoparticles were found in only 7% of hepatocytes. Depletion of Kupffer cells by clodronate liposomes increased nanoparticle retention in liver sinusoidal endothelial cells and hepatic stellate cells, but not in hepatocytes. It is importantly suggested that studies of drug-loaded nanoparticle delivery to the liver have to demonstrate not only uptake of nanoparticles by the target cell type but also non-uptake by other cell types to assess their effect as well as ensure their safety.

DOI： 10.1016/j.nano.2016.01.013

Pancreatic α-Amylase Controls Glucose Assimilation by Duodenal Retrieval through N-Glycan-specific Binding, Endocytosis, and Degradation 査読国際誌

Kimie Date, Ayano Satoh, Kaoruko Iida, Haruko Ogawa

Journal of Biological Chemistry 290 ( 28 ) 17439 - 17450 2015年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

α-Amylase, a major pancreatic protein and starch hydrolase, is essential for energy acquisition. Mammalian pancreatic α-amylase binds specifically to glycoprotein N-glycans in the brush-border membrane to activate starch digestion, whereas it significantly inhibits glucose uptake by Na(+)/glucose cotransporter 1 (SGLT1) at high concentrations (Asanuma-Date, K., Hirano, Y., Le, N., Sano, K., Kawasaki, N., Hashii, N., Hiruta, Y., Nakayama, K., Umemura, M., Ishikawa, K., Sakagami, H., and Ogawa, H. (2012) Functional regulation of sugar assimilation by N-glycan-specific interaction of pancreatic α-amylase with glycoproteins of duodenal brush border membrane. J. Biol. Chem. 287, 23104-23118). However, how the inhibition is stopped was unknown. Here, we show a new mechanism for the regulation of intestinal glucose absorption. Immunohistochemistry revealed that α-amylase in the duodena of non-fasted, but not fasted, pigs was internalized from the pancreatic fluid and immunostained. We demonstrated that after N-glycan binding, pancreatic α-amylase underwent internalization into lysosomes in a process that was inhibited by α-mannoside. The internalized α-amylase was degraded, showing low enzymatic activity and molecular weight at the basolateral membrane. In a human intestinal Caco-2 cell line, Alexa Fluor 488-labeled pancreatic α-amylase bound to the cytomembrane was transported to lysosomes through the endocytic pathway and then disappeared, suggesting degradation. Our findings indicate that N-glycan recognition by α-amylase protects enterocytes against a sudden increase in glucose concentration and restores glucose uptake by gradual internalization, which homeostatically controls the postprandial blood glucose level. The internalization of α-amylase may also enhance the supply of amino acids required for the high turnover of small intestine epithelial cells. This study provides novel and significant insights into the control of blood sugar during the absorption stage in the intestine.

DOI： 10.1074/jbc.m114.594937

The Golgin Tether Giantin Regulates the Secretory Pathway by Controlling Stack Organization within Golgi Apparatus 査読国際誌

Mayuko Koreishi, Thomas J. Gniadek, Sidney Yu, Junko Masuda, Yasuko Honjo, Ayano Satoh

PLoS ONE 8 ( 3 ) e59821 - e59821 2013年3月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Public Library of Science (PLoS)

Golgins are coiled-coil proteins that play a key role in the regulation of Golgi architecture and function. Giantin, the largest golgin in mammals, forms a complex with p115, rab1, GM130, and soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), thereby facilitating vesicle tethering and fusion processes around the Golgi apparatus. Treatment with the microtubule destabilizing drug nocodazole transforms the Golgi ribbon into individual Golgi stacks. Here we show that siRNA-mediated depletion of giantin resulted in more dispersed Golgi stacks after nocodazole treatment than by control treatment, without changing the average cisternal length. Furthermore, depletion of giantin caused an increase in cargo transport that was associated with altered cell surface protein glycosylation. Drosophila S2 cells are known to have dispersed Golgi stacks and no giantin homolog. The exogenous expression of mammalian giantin cDNA in S2 cells resulted in clustered Golgi stacks, similar to the Golgi ribbon in mammalian cells. These results suggest that the spatial organization of the Golgi ribbon is mediated by giantin, which also plays a role in cargo transport and sugar modifications.

DOI： 10.1371/journal.pone.0059821

Stabilization of Actin Bundles by a Dynamin 1/Cortactin Ring Complex Is Necessary for Growth Cone Filopodia 査読国際誌

Hiroshi Yamada, Tadashi Abe, Ayano Satoh, Nana Okazaki, Shota Tago, Kinue Kobayashi, Yumi Yoshida, Yoshiya Oda, Masami Watanabe, Kazuhito Tomizawa, Hideki Matsui, Kohji Takei

The Journal of Neuroscience 33 ( 10 ) 4514 - 4526 2013年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Society for Neuroscience

Dynamin GTPase, a key molecule in endocytosis, mechanically severs the invaginated membrane upon GTP hydrolysis. Dynamin functions also in regulating actin cytoskeleton, but the mechanisms are yet to be defined. Here we show that dynamin 1, a neuronal isoform of dynamin, and cortactin form ring complexes, which twine around F-actin bundles and stabilize them. By negative-staining EM, dynamin 1–cortactin complexes appeared as “open” or “closed” rings depending on guanine nucleotide conditions. By pyrene actin assembly assay, dynamin 1 stimulated actin assembly in mouse brain cytosol.In vitroincubation of F-actin with both dynamin 1 and cortactin led to the formation of long and thick actin bundles, on which dynamin 1 and cortactin were periodically colocalized in puncta. A depolymerization assay revealed that dynamin 1 and cortactin increased the stability of actin bundles, most prominently in the presence of GTP. In rat cortical neurons and human neuroblastoma cell line, SH-SY5Y, both dynamin 1 and cortactin localized on actin filaments and the bundles at growth cone filopodia as revealed by immunoelectron microscopy. In SH-SY5Y cell, acute inhibition of dynamin 1 by application of dynamin inhibitor led to growth cone collapse. Cortactin knockdown also reduced growth cone filopodia. Together, our results strongly suggest that dynamin 1 and cortactin ring complex mechanically stabilizes F-actin bundles in growth cone filopodia. Thus, the GTPase-dependent mechanochemical enzyme property of dynamin is commonly used both in endocytosis and regulation of F-actin bundles by a dynamin 1–cortactin complex.

DOI： 10.1523/jneurosci.2762-12.2013

Absence of Nogo-B (Reticulon 4B) Facilitates Hepatic Stellate Cell Apoptosis and Diminishes Hepatic Fibrosis in Mice 査読国際誌

Keitaro Tashiro, Ayano Satoh, Teruo Utsumi, Chuhan Chung, Yasuko Iwakiri

The American Journal of Pathology 182 ( 3 ) 786 - 795 2013年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

Nogo-B (reticulon 4B) accentuates hepatic fibrosis and cirrhosis, but the mechanism remains unclear. The aim of this study was to identify the role of Nogo-B in hepatic stellate cell (HSC) apoptosis in cirrhotic livers. Cirrhosis was generated by carbon tetrachloride inhalation in wild-type (WT) and Nogo-A/B knockout (Nogo-B KO) mice. HSCs were isolated from WT and Nogo-B KO mice and cultured for activation and transformation to myofibroblasts (MF-HSCs). Human hepatic stellate cells (LX2 cells) were used to assess apoptotic responses of activated HSCs after silencing or overexpressing Nogo-B. Livers from cirrhotic Nogo-B KO mice showed significantly reduced fibrosis (P < 0.05) compared with WT mice. Apoptotic cells were more prominent in fibrotic areas of cirrhotic Nogo-B KO livers. Nogo-B KO MF-HSCs showed significantly increased levels of apoptotic markers, cleaved poly (ADP-ribose) polymerase, and caspase-3 and -8 (P < 0.05) compared with WT MF-HSCs in response to staurosporine. Treatment with tunicamycin, an endoplasmic reticulum stress inducer, increased cleaved caspase-3 and -8 levels in Nogo-B KO MF-HSCs compared with WT MF-HSCs (P < 0.01). In LX2 cells, Nogo-B knockdown enhanced apoptosis in response to staurosporine, whereas Nogo-B overexpression inhibited apoptosis. The absence of Nogo-B enhances apoptosis of HSCs in experimental cirrhosis. Selective blockade of Nogo-B in HSCs may represent a potential therapeutic strategy to mitigate liver fibrosis.

DOI： 10.1016/j.ajpath.2012.11.032

CK2 Phosphorylates Sec31 and Regulates ER-To-Golgi Trafficking 国際誌

Mayuko Koreishi, Sidney Yu, Mayumi Oda, Yasuko Honjo, Ayano Satoh

PLoS ONE 8 ( 1 ) e54382 - e54382 2013年1月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Public Library of Science (PLoS)

Protein export from the endoplasmic reticulum (ER) is an initial and rate-limiting step of molecular trafficking and secretion. This is mediated by coat protein II (COPII)-coated vesicles, whose formation requires small GTPase Sar1 and 6 Sec proteins including Sec23 and Sec31. Sec31 is a component of the outer layer of COPII coat and has been identified as a phosphoprotein. The initiation and promotion of COPII vesicle formation is regulated by Sar1; however, the mechanism regulating the completion of COPII vesicle formation followed by vesicle release is largely unknown. Hypothesizing that the Sec31 phosphorylation may be such a mechanism, we identified phosphorylation sites in the middle linker region of Sec31. Sec31 phosphorylation appeared to decrease its association with ER membranes and Sec23. Non-phosphorylatable mutant of Sec31 stayed longer at ER exit sites and bound more strongly to Sec23. We also found that CK2 is one of the kinases responsible for Sec31 phosphorylation because CK2 knockdown decreased Sec31 phosphorylation, whereas CK2 overexpression increased Sec31 phosphorylation. Furthermore, CK2 knockdown increased affinity of Sec31 for Sec23 and inhibited ER-to-Golgi trafficking. These results suggest that Sec31 phosphorylation by CK2 controls the duration of COPII vesicle formation, which regulates ER-to-Golgi trafficking.

DOI： 10.1371/journal.pone.0054382

TRAPPC9 Mediates the Interaction between p150Glued and COPII Vesicles at the Target Membrane 査読国際誌

Min Zong, Ayano Satoh, Mei Kuen Yu, Ka Yu Siu, Wing Yan Ng, Hsiao Chang Chan, Julian A. Tanner, Sidney Yu

PLoS ONE 7 ( 1 ) e29995 - e29995 2012年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Public Library of Science (PLoS)

BACKGROUND: The transport of endoplasmic reticulum (ER)-derived COPII vesicles toward the ER-Golgi intermediate compartment (ERGIC) requires cytoplasmic dynein and is dependent on microtubules. p150(Glued), a subunit of dynactin, has been implicated in the transport of COPII vesicles via its interaction with COPII coat components Sec23 and Sec24. However, whether and how COPII vesicle tether, TRAPP (Transport protein particle), plays a role in the interaction between COPII vesicles and microtubules is currently unknown. PRINCIPLE FINDINGS: We address the functional relationship between COPII tether TRAPP and dynactin. Overexpressed TRAPP subunits interfered with microtubule architecture by competing p150(Glued) away from the MTOC. TRAPP subunit TRAPPC9 bound directly to p150(Glued) via the same carboxyl terminal domain of p150(Glued) that binds Sec23 and Sec24. TRAPPC9 also inhibited the interaction between p150(Glued) and Sec23/Sec24 both in vitro and in vivo, suggesting that TRAPPC9 serves to uncouple p150(Glued) from the COPII coat, and to relay the vesicle-dynactin interaction at the target membrane. CONCLUSIONS: These findings provide a new perspective on the function of TRAPP as an adaptor between the ERGIC membrane and dynactin. By preserving the connection between dynactin and the tethered and/or fused vesicles, TRAPP allows nascent ERGIC to continue the movement along the microtubules as they mature into the cis-Golgi.

DOI： 10.1371/journal.pone.0029995

Abnormal Trafficking and Degradation of TLR4 Underlie the Elevated Inflammatory Response in Cystic Fibrosis 査読国際誌

Emanuela M. Bruscia, Ping-Xia Zhang, Ayano Satoh, Christina Caputo, Ruslan Medzhitov, Ambika Shenoy, Marie E. Egan, Diane S. Krause

The Journal of Immunology 186 ( 12 ) 6990 - 6998 2011年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：The American Association of Immunologists

Abstract

Morbidity and mortality in cystic fibrosis (CF) are due not only to abnormal epithelial cell function, but also to an abnormal immune response. We have shown previously that macrophages lacking CF transmembrane conductance regulator (CFTR), the gene mutated in CF, contribute significantly to the hyperinflammatory response observed in CF. In this study, we show that lack of functional CFTR in murine macrophages causes abnormal TLR4 subcellular localization. Upon LPS stimulation, CFTR macrophages have prolonged TLR4 retention in the early endosome and reduced translocation into the lysosomal compartment. This abnormal TLR4 trafficking leads to increased LPS-induced activation of the NF-κB, MAPK, and IFN regulatory factor-3 pathways and decreased TLR4 degradation, which affects downregulation of the proinflammatory state. In addition to primary murine cells, mononuclear cells isolated from CF patients demonstrate similar defects in response to LPS. Moreover, specific inhibition of CFTR function induces abnormal TLR4 trafficking and enhances the inflammatory response of wild-type murine cells to LPS. Thus, functional CFTR in macrophages influences TLR4 spatial and temporal localization and perturbs LPS-mediated signaling in both murine CF models and patients with CF.

DOI： 10.4049/jimmunol.1100396

Detection of in situ cleaved p115 with the cut specific antibodies in rapid protein inactivation system by tobacco etch viral protease cleavage 査読

Koreyoshi, Mayuko, Honjo, Yasuko, Satoh, Ayano

Antibody Technology Journal 2011 ( 1 ) 5 - 11 2011年

mTrs130 Is a Component of a Mammalian TRAPPII Complex, a Rab1 GEF That Binds to COPI-coated Vesicles 査読国際誌

Akinori Yamasaki, Shekar Menon, Sidney Yu, Jemima Barrowman, Timo Meerloo, Viola Oorschot, Judith Klumperman, Ayano Satoh, Susan Ferro-Novick

Molecular Biology of the Cell 20 ( 19 ) 4205 - 4215 2009年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：American Society for Cell Biology (ASCB)

The GTPase Rab1 regulates endoplasmic reticulum-Golgi and early Golgi traffic. The guanine nucleotide exchange factor (GEF) or factors that activate Rab1 at these stages of the secretory pathway are currently unknown. Trs130p is a subunit of the yeast TRAPPII (transport protein particle II) complex, a multisubunit tethering complex that is a GEF for the Rab1 homologue Ypt1p. Here, we show that mammalian Trs130 (mTrs130) is a component of an analogous TRAPP complex in mammalian cells, and we describe for the first time the role that this complex plays in membrane traffic. mTRAPPII is enriched on COPI (Coat Protein I)-coated vesicles and buds, but not Golgi cisternae, and it specifically activates Rab1. In addition, we find that mTRAPPII binds to γ1COP, a COPI coat adaptor subunit. The depletion of mTrs130 by short hairpin RNA leads to an increase of vesicles in the vicinity of the Golgi and the accumulation of cargo in an early Golgi compartment. We propose that mTRAPPII is a Rab1 GEF that tethers COPI-coated vesicles to early Golgi membranes.

DOI： 10.1091/mbc.e09-05-0387

Following the Fate In Vivo of COPI Vesicles Generated In Vitro 査読国際誌

Christoph Rutz, Ayano Satoh, Paolo Ronchi, Britta Brügger, Graham Warren, Felix T. Wieland

Traffic 10 ( 8 ) 994 - 1005 2009年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

COPI vesicles are a class of transport carriers that function in the early secretory pathway. Their fate and function are still controversial. This includes their contribution to bidirectional transport within the Golgi apparatus and their role during cell division. Here we describe a method that should address several open questions about the fate and function of COPI vesicles in vivo. To this end, fluorescently labeled COPI vesicles were generated in vitro from isolated rat liver Golgi membranes, labeled with the fluorescent dyes Alexa‐488 or Alexa‐568. These vesicles appeared to be active and colocalized with endogenous Golgi membranes within 30 min after microinjection into mammalian cells. The COPI vesicle‐derived labeled membrane proteins could be classified into two types that behaved like endogenous proteins after Brefeldin A treatment.

DOI： 10.1111/j.1600-0854.2009.00934.x

A PDZ‐Binding Motif Controls Basolateral Targeting of Syndecan‐1 Along the Biosynthetic Pathway in Polarized Epithelial Cells 査読国際誌

Sandra Maday, Eric Anderson, Henry C. Chang, James Shorter, Ayano Satoh, Jeff Sfakianos, Heike Fölsch, James M. Anderson, Zenta Walther, Ira Mellman

Traffic 9 ( 11 ) 1915 - 1924 2008年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

The cell surface proteoglycan, syndecan‐1, is essential for normal epithelial morphology and function. Syndecan‐1 is selectively localized to the basolateral domain of polarized epithelial cells and interacts with cytosolic PDZ (PSD‐95, discs large, ZO‐1) domain‐containing proteins. Here, we show that the polarity of syndecan‐1 is determined by its type II PDZ‐binding motif. Mutations within the PDZ‐binding motif lead to the mislocalization of syndecan‐1 to the apical surface. In contrast to previous examples, however, PDZ‐binding motif‐dependent polarity is not determined by retention at the basolateral surface but rather by polarized sorting prior to syndecan‐1’s arrival at the plasma membrane. Although none of the four known PDZ‐binding partners of syndecan‐1 appears to control basolateral localization, our results show that the PDZ‐binding motif of syndecan‐1 is decoded along the biosynthetic pathway establishing a potential role for PDZ‐mediated interactions in polarized sorting.

DOI： 10.1111/j.1600-0854.2008.00805.x

In Situ Cleavage of the Acidic Domain from the p115 Tether Inhibits Exocytic Transport 査読国際誌

Ayano Satoh, Graham Warren

Traffic 9 ( 9 ) 1522 - 1529 2008年8月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

Golgins are coiled‐coil proteins involved in Golgi architecture and function. A complex of golgins (p115, GM130 and giantin), together with the rab1 guanosine triphosphatase and cis Golgi SNAREs, helps to mediate fusion processes at the entry face of the Golgi apparatus. The C‐terminal acidic domain of p115 binds specifically to GM130 and giantin. However, deletion of this domain in vivo appears to have no effect on exocytic transport when using an RNA interference depletion/rescue approach (Puthenveedu MA, Linstedt AD. Gene replacement reveals that p115/SNARE interactions are essential for Golgi biogenesis. Proc Natl Acad Sci U S A 2004;101:1253–1256). In this study, we have used a different approach introducing a tobacco etch virus (tev) protease cleavage site into p115 so that the C‐terminal domain can be rapidly and specifically released in vivo by microinjection of the tev protease. The results show that cleavage inhibits exocytic transport to the cell surface.

DOI： 10.1111/j.1600-0854.2008.00783.x

Ankyrin Repeat Proteins Comprise a Diverse Family of Bacterial Type IV Effectors 査読国際誌

Xiaoxiao Pan, Anja Lührmann, Ayano Satoh, Michelle A. Laskowski-Arce, Craig R. Roy

Science 320 ( 5883 ) 1651 - 1654 2008年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：American Association for the Advancement of Science (AAAS)

Specialized secretion systems are used by many bacteria to deliver effector proteins into host cells that can either mimic or disrupt the function of eukaryotic factors. We found that the intracellular pathogens Legionella pneumophila and Coxiella burnetii use a type IV secretion system to deliver into eukaryotic cells a large number of different bacterial proteins containing ankyrin repeat homology domains called Anks. The L. pneumophila AnkX protein prevented microtubule-dependent vesicular transport to interfere with fusion of the L. pneumophila -containing vacuole with late endosomes after infection of macrophages, which demonstrates that Ank proteins have effector functions important for bacterial infection of eukaryotic host cells.

DOI： 10.1126/science.1158160

Nitric oxide synthase generates nitric oxide locally to regulate compartmentalized protein S-nitrosylation and protein trafficking 査読国際誌

Yasuko Iwakiri, Ayano Satoh, Suvro Chatterjee, Derek K. Toomre, Cecile M. Chalouni, David Fulton, Roberto J. Groszmann, Vijay H. Shah, William C. Sessa

Proceedings of the National Academy of Sciences 103 ( 52 ) 19777 - 19782 2006年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Proceedings of the National Academy of Sciences

Nitric oxide (NO) is a highly diffusible and short-lived physiological messenger. Despite its diffusible nature, NO modifies thiol groups of specific cysteine residues in target proteins and alters protein function via S-nitrosylation. Although intracellular S-nitrosylation is a specific posttranslational modification, the defined localization of an NO source (nitric oxide synthase, NOS) with protein S-nitrosylation has never been directly demonstrated. Endothelial NOS (eNOS) is localized mainly on the Golgi apparatus and in plasma membrane caveolae. Here, we show by using eNOS targeted to either the Golgi or the nucleus that S-nitrosylation is concentrated at the primary site of eNOS localization. Furthermore, localization of eNOS on the Golgi enhances overall Golgi protein S-nitrosylation, the specific S-nitrosylation of N -ethylmaleimide-sensitive factor and reduces the speed of protein transport from the endoplasmic reticulum to the plasma membrane in a reversible manner. These data indicate that local NOS action generates organelle-specific protein S-nitrosylation reactions that can regulate intracellular transport processes.

DOI： 10.1073/pnas.0605907103

mBet3p is required for homotypic COPII vesicle tethering in mammalian cells 査読国際誌

Sidney Yu, Ayano Satoh, Marc Pypaert, Karl Mullen, Jesse C. Hay, Susan Ferro-Novick

The Journal of Cell Biology 174 ( 3 ) 359 - 368 2006年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Rockefeller University Press

TRAPPI is a large complex that mediates the tethering of COPII vesicles to the Golgi (heterotypic tethering) in the yeast Saccharomyces cerevisiae. In mammalian cells, COPII vesicles derived from the transitional endoplasmic reticulum (tER) do not tether directly to the Golgi, instead, they appear to tether to each other (homotypic tethering) to form vesicular tubular clusters (VTCs). We show that mammalian Bet3p (mBet3p), which is the most highly conserved TRAPP subunit, resides on the tER and adjacent VTCs. The inactivation of mBet3p results in the accumulation of cargo in membranes that colocalize with the COPII coat. Furthermore, using an assay that reconstitutes VTC biogenesis in vitro, we demonstrate that mBet3p is required for the tethering and fusion of COPII vesicles to each other. Consistent with the proposal that mBet3p is required for VTC biogenesis, we find that ERGIC-53 (VTC marker) and Golgi architecture are disrupted in siRNA-treated mBet3p-depleted cells. These findings imply that the TRAPPI complex is essential for VTC biogenesis.

DOI： 10.1083/jcb.200603044

A Cryptic Rab1-binding Site in the p115 TetheringProtein 査読国際誌

Matthew Beard, Ayano Satoh, James Shorter, Graham Warren

Journal of Biological Chemistry 280 ( 27 ) 25840 - 25848 2005年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

Small GTPases and coiled-coil proteins of the golgin family help to tether COPI vesicles to Golgi membranes. At the cis-side of the Golgi, the Rab1 GTPase binds directly to each of three coiled-coil proteins: p115, GM130, and as now shown, Giantin. Rab1 binds to a coiled-coil region within the tail domain of p115 and this binding is inhibited by the C-terminal, acidic domain of p115. Furthermore, GM130 and Giantin bind to the acidic domain of p115 and stimulate p115 binding to Rab1, suggesting that p115 binding to Rab1 is regulated. Regulation of this interaction by proteins such as GM130 and Giantin may control the membrane recruitment of p115 by Rab1.

DOI： 10.1074/jbc.m503925200

Golgin Tethers Define Subpopulations of COPI Vesicles 査読国際誌

Jörg Malsam, Ayano Satoh, Laurence Pelletier, Graham Warren

Science 307 ( 5712 ) 1095 - 1098 2005年2月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：American Association for the Advancement of Science (AAAS)

Coiled-coil proteins of the golgin family have been implicated in intra-Golgi transport through tethering coat protein complex I (COPI) vesicles. The p115-golgin tether is the best studied, and here we characterize the golgin-84–CASP tether. The vesicles bound by this tether were strikingly different from those bound by the p115-golgin tether in that they lacked members of the p24 family of putative cargo receptors and contained enzymes instead of anterograde cargo. Microinjected golgin-84 or CASP also inhibited Golgi-enzyme transport to the endoplasmic reticulum, further implicating this tether in retrograde transport. These and other golgins may modulate the flow patterns within the Golgi stack.

DOI： 10.1126/science.1108061

Mapping the Functional Domains of the Golgi Stacking Factor GRASP65 査読国際誌

Yanzhuang Wang, Ayano Satoh, Graham Warren

Journal of Biological Chemistry 280 ( 6 ) 4921 - 4928 2005年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

The Golgi reassembly stacking protein (GRASP) family has been implicated in the stacking of Golgi cisternae and the regulation of Golgi disassembly/reassembly during mitosis in mammalian cells. GRASP65 is a dimer that can directly link adjacent surfaces through trans-oligomerization in a mitotically regulated manner. Here we show that the N-terminal GRASP domain (amino acids 1-201) is both necessary and sufficient for dimerization and trans-oligomerization but is not mitotically regulated. The C-terminal serine/proline-rich domain (amino acids 202-446) cannot dimerize nor can it link adjacent surfaces. It does, however, confer mitotic regulation on the GRASP domain through multiple sites phosphorylated by the mitotic kinases, cdc2/B1, and the polo-like kinase. Transient expression corroborated these results by showing that the GRASP domain alone inhibited mitotic fragmentation of the Golgi apparatus.

DOI： 10.1074/jbc.m412407200

Tethering Assays for COPI Vesicles Mediated by Golgins 査読国際誌

Ayano Satoh, Jörg Malsam, Graham Warren

Methods in Enzymology 404 125 - 134 2005年

担当区分：筆頭著者記述言語：英語掲載種別：論文集(書籍)内論文出版者・発行元：Elsevier

A method is described that allows the attachment of COPI vesicles and Golgi membranes to glass slides that can then be analyzed using electron microscopy (EM) and immuno-EM methods. Subpopulations of COPI vesicles can be bound selectively using recombinant golgins. Alternatively, COPI vesicles can be attached to prebound Golgi membranes. Marking these vesicles selectively with biotin allows their site of attachment to be identified.

DOI： 10.1016/s0076-6879(05)04013-9

Preparation and Characterization of Recombinant Golgin Tethers 査読国際誌

Ayano Satoh, Matthew Beard, Graham Warren

Methods in Enzymology 404 279 - 296 2005年

担当区分：筆頭著者記述言語：英語掲載種別：論文集(書籍)内論文出版者・発行元：Elsevier

Golgin tethers are integral or peripheral Golgi proteins with predicted coiled-coil domains and many are known to interact directly with small GTPases of the Ypt/Rab or Arl families. Here we describe the preparation of recombinant golgins: GM130, p115 (and truncations thereof), the N-terminal fragment of giantin, CASP, and golgin-84.

DOI： 10.1016/s0076-6879(05)04026-7

VCIP135 acts as a deubiquitinating enzyme during p97–p47-mediated reassembly of mitotic Golgi fragments 査読国際誌

Yanzhuang Wang, Ayano Satoh, Graham Warren, Hemmo H. Meyer

The Journal of Cell Biology 164 ( 7 ) 973 - 978 2004年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Rockefeller University Press

The AAA-ATPase p97/Cdc48 functions in different cellular pathways using distinct sets of adapters and other cofactors. Together with its adaptor Ufd1–Npl4, it extracts ubiquitylated substrates from the membrane for subsequent delivery to the proteasome during ER-associated degradation. Together with its adaptor p47, on the other hand, it regulates several membrane fusion events, including reassembly of Golgi cisternae after mitosis. The finding of a ubiquitin-binding domain in p47 raises the question as to whether the ubiquitin–proteasome system is also involved in membrane fusion events. Here, we show that p97–p47-mediated reassembly of Golgi cisternae requires ubiquitin, but is not dependent on proteasome-mediated proteolysis. Instead, it requires the deubiquitinating activity of one of its cofactors, VCIP135, which reverses a ubiquitylation event that occurs during mitotic disassembly. Together, these data reveal a cycle of ubiquitylation and deubiquitination that regulates Golgi membrane dynamics during mitosis. Furthermore, they represent the first evidence for a proteasome-independent function of p97/Cdc48.

DOI： 10.1083/jcb.200401010

Golgin‐84 is a rab1 Binding Partner Involved in Golgi Structure 査読国際誌

Ayano Satoh, Yanzhuang Wang, Jörg Malsam, Matthew B. Beard, Graham Warren

Traffic 4 ( 3 ) 153 - 161 2003年3月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

Members of the golgin family of coiled‐coil proteins have been implicated in the tethering of vesicles to Golgi membranes and cisternal membranes to each other. Many also bind to rab GTPases. Golgin‐84 is a membrane‐anchored golgin that we now show binds preferentially to the GTP form of the rab1 GTPase. It is also present throughout the Golgi stack by immuno‐EM. Antibodies to golgin‐84 inhibit stacking of cisternal membranes in a cell‐free assay for Golgi reassembly, whereas the cytoplasmic domain of golgin‐84 stimulates stacking and increases the length of re‐assembled stacks. Transient expression of golgin‐84 in NRK cells helps prevent the disassembly of the Golgi apparatus normally triggered by treatment with brefeldin A. Together these data suggest that golgin‐84 is involved in generating and maintaining the architecture of the Golgi apparatus.

DOI： 10.1034/j.1600-0854.2003.00103.x

Fraglide-1 from traditional Chinese aromatic vinegar: A natural AhR antagonist for atopic dermatitis 国際誌

Kosuke Kato, Miki Akamatsu, Saya Kakimaru, Mayuko Koreishi, Masahiro Takagi, Masahiro Miyashita, Yoshiyuki Murata, Yoshimasa Nakamura, Ayano Satoh, Yoshio Tsujino

Food and Chemical Toxicology 197 115301 - 115301 2025年3月

記述言語：英語掲載種別：研究論文（学術雑誌）

Traditional Chinese Zhenjiang aromatic vinegar (Kozu) contains Fraglide-1 (FG1), a bioactive lactone with demonstrated peroxisome proliferator-activated receptor gamma (PPARγ) agonist and antioxidant activities. This study explored FG1's novel ability to antagonize the aryl hydrocarbon receptor (AhR) signaling pathway, which regulates artemin expression and contributes to itching and inflammation in atopic dermatitis. Through molecular docking simulations and cell-based assays in human keratinocytes, we demonstrated FG1's potent antagonistic activity against AhR signaling. FG1 effectively suppressed FICZ-induced inflammatory responses, including artemin expression, with potency (half maximal inhibitory concentration, IC50 = 5.1 μM) comparable to the synthetic antagonist StemRegenin 1 (SR1) while demonstrating a superior safety profile (median lethal concentration, LC50 > 100 μM vs. 27.5 μM for SR1). These findings expand our understanding of bioactive compounds from traditional fermented foods and their regulatory effects on AhR signaling, providing a foundation for future studies on FG1's role in modulating skin inflammation.

DOI： 10.1016/j.fct.2025.115301

Resveratrol, a food-derived polyphenol, promotes Melanosomal degradation in skin fibroblasts through coordinated activation of autophagy, lysosomal, and antioxidant pathways

Saki Okamoto, Saya Kakimaru, Mayuko Koreishi, Mika Sakamoto, Yoshimasa Nakamura, Hideya Ando, Yoshio Tsujino, Ayano Satoh

Journal of Functional Foods 2025年2月

PTPA localized in the Golgi apparatus plays an important role in osteoblast differentiation.

Zheng X, Weng Y, Satoh A, Tanai A, Ikegame M, Kimura K, Yoshitani N, Xie X, Okamura H

Biochemical and biophysical research communications 748 151329 - 151329 2025年1月

掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

Regulatory subunits of protein phosphatase 2A (PP2A) define the substrate and functional specificity of the PP2A holoenzyme within specific organelles. While PP2A regulates osteoblast differentiation, the roles and localization of its regulatory subunits in osteoblasts remain unclear. Here, we identified PTPA, a PP2A regulatory protein, predominantly localized to the Golgi apparatus, closely overlapping with the Golgi marker Giantin. Disruption of the Golgi structure by Brefeldin A caused PTPA to disperse into the cytoplasm. PTPA overexpression inhibited osteoblast differentiation by downregulating key transcriptional regulators. The Golgi-specific localization of PTPA suggests it may influence bone-related protein secretion and maintain Golgi integrity. These findings highlight the critical role of PTPA in osteoblast differentiation and its association with the Golgi apparatus.

DOI： 10.1016/j.bbrc.2025.151329

Evaluation of quercetin as a potential cytoprotector against acetaldehyde using the cultured hepatocyte model with aldehyde dehydrogenase isozyme deficiency. 査読国際誌

Yuhang Xu, Takeshi Sawamoto, Ruitong Sun, Aki Ishikura, Shintaro Munemasa, Yoshiyuki Murata, Ayano Satoh, Akiko Matsumoto, Toshiyuki Nakamura, Yoshimasa Nakamura

Bioscience, biotechnology, and biochemistry 88 ( 10 ) 1199 - 1202 2024年9月

記述言語：英語掲載種別：研究論文（学術雑誌）

Protective effect of quercetin against acetaldehyde was evaluated using the cultured hepatocyte models with aldehyde dehydrogenase (ALDH) isozyme deficiency (aldh2-kd and aldh1a1-kd). The quercetin-induced cytoprotection against acetaldehyde in the ALDH1A1-deficient mutant (aldh1a1-kd) was weaker than that in the wild type. Furthermore, quercetin did not enhance the ALDH activity in aldh1a1-kd cells, suggesting that ALDH1A1 is involved in quercetin-induced cytoprotection.

DOI： 10.1093/bbb/zbae100

Development of a novel AAK1 inhibitor via Kinobeads-based screening. 査読国際誌

Akari Yoshida, Satomi Ohtsuka, Fumiya Matsumoto, Tomoyuki Miyagawa, Rei Okino, Yumeya Ikeda, Natsume Tada, Akira Gotoh, Masaki Magari, Naoya Hatano, Ryo Morishita, Ayano Satoh, Yukinari Sunatsuki, Ulf J Nilsson, Teruhiko Ishikawa, Hiroshi Tokumitsu

Scientific reports 14 ( 1 ) 6723 - 6723 2024年3月

記述言語：英語掲載種別：研究論文（学術雑誌）

A chemical proteomics approach using Ca2+/calmodulin-dependent protein kinase kinase (CaMKK) inhibitor-immobilized sepharose (TIM-063-Kinobeads) identified main targets such as CaMKKα/1 and β/2, and potential off-target kinases, including AP2-associated protein kinase 1 (AAK1), as TIM-063 interactants. Because TIM-063 interacted with the AAK1 catalytic domain and inhibited its enzymatic activity moderately (IC50 = 8.51 µM), we attempted to identify potential AAK1 inhibitors from TIM-063-derivatives and found a novel AAK1 inhibitor, TIM-098a (11-amino-2-hydroxy-7H-benzo[de]benzo[4,5]imidazo[2,1-a]isoquinolin-7-one) which is more potent (IC50 = 0.24 µM) than TIM-063 without any inhibitory activity against CaMKK isoforms and a relative AAK1-selectivity among the Numb-associated kinases family. TIM-098a could inhibit AAK1 activity in transfected cultured cells (IC50 = 0.87 µM), indicating cell-membrane permeability of the compound. Overexpression of AAK1 in HeLa cells significantly reduced the number of early endosomes, which was blocked by treatment with 10 µM TIM-098a. These results indicate TIM-063-Kinobeads-based chemical proteomics is efficient for identifying off-target kinases and re-evaluating the kinase inhibitor (TIM-063), leading to the successful development of a novel inhibitory compound (TIM-098a) for AAK1, which could be a molecular probe for AAK1. TIM-098a may be a promising lead compound for a more potent, selective and therapeutically useful AAK1 inhibitor.

DOI： 10.1038/s41598-024-57051-9

Aromatic oil from lavender as an atopic dermatitis suppressant 査読国際誌

Haruna Sato, Kosuke Kato, Mayuko Koreishi, Yoshimasa Nakamura, Yoshio Tsujino, Ayano Satoh

PLOS ONE 19 ( 1 ) e0296408 2024年1月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

In atopic dermatitis (AD), nerves are abnormally stretched near the surface of the skin, making it sensitive to itching. Expression of neurotrophic factor Artemin (ARTN) involved in such nerve stretching is induced by the xenobiotic response (XRE) to air pollutants and UV radiation products. Therefore, AD can be monitored by the XRE response. Previously, we established a human keratinocyte cell line stably expressing a NanoLuc reporter gene downstream of XRE. We found that 6-formylindolo[3,2-b]carbazole (FICZ), a tryptophan metabolite and known inducer of the XRE, increased reporter and Artemin mRNA expression, indicating that FICZ-treated cells could be a model for AD. Lavender essential oil has been used in folk medicine to treat AD, but the scientific basis for its use is unclear. In the present study, we investigated the efficacy of lavender essential oil and its major components, linalyl acetate and linalool, to suppress AD and sensitize skin using the established AD model cell line, and keratinocyte and dendritic cell activation assays. Our results indicated that lavender essential oil from L. angustifolia and linalyl acetate exerted a strong AD inhibitory effect and almost no skin sensitization. Our model is useful in that it can circumvent the practice of using animal studies to evaluate AD medicines.

DOI： 10.1371/journal.pone.0296408

DDS-type near-infrared light absorber enables deeper lesion treatment in laser photothermal therapy while avoiding damage to surrounding organs. 査読国際誌

Masataka Takahashi, Jun Fujishiro, Shinsuke Nomura, Manabu Harada, Akinari Hinoki, Masashi Arake, Eiichi Ozeki, Isao Hara, Ayano Satoh, Takahisa Tainaka, Hiro-O Uchida, Yuji Morimoto

Frontiers in bioengineering and biotechnology 12 1444107 - 1444107 2024年

記述言語：英語掲載種別：研究論文（学術雑誌）

The efficacy of drug delivery system (DDS)-type near-infrared (NIR) absorbing agents in enhancing laser photothermal therapy is widely acknowledged. Despite the acknowledged efficacy, the therapeutic advantages of photothermal therapy using DDS-type NIR-absorbing agents over simple photothermal therapy without such agents have not been fully elucidated. This study was designed to investigate two primary objectives: firstly, the ability of DDS-type NIR-absorbing agents to induce cell death at greater depths within tumors, and secondly, their capacity to minimize collateral damage to adjacent healthy organs. To investigate these objectives, we employed a combination of indocyanine green lactosome-a DDS-type NIR-absorbing agent-and a precision-controlled laser hyperthermia system. An orthotopic neuroblastoma tumor model was used to closely simulate clinical conditions. The findings revealed that photothermal therapy using the DDS-type NIR-absorbing agent not only facilitates deeper penetration of cell death within tumors but also significantly mitigates thermal damage to surrounding healthy tissues, when compared to simple phototherapy without the agent. Furthermore, the combined treatment significantly prolonged the survival periods of the animals involved. This study is the first to analyze these therapeutic efficacies using quantitative data from an orthotopic tumor animal model and substantiated the potential of DDS-type NIR-absorbing agents to deepen the therapeutic impact of photothermal therapy while safeguarding vital organs, thereby enhancing overall treatment outcomes.

DOI： 10.3389/fbioe.2024.1444107

Cancer stem cells as the source of tumor associated myoepithelial cells in the tumor microenvironment developing ductal carcinoma in situ 査読国際誌

Said M. Afify, Ghmkin Hassan, Maram H. Zahra, Hend M. Nawara, Hagar A. Abu Quora, Amira Osman, Hager Mansour, Kazuki Kumon, Akimasa Seno, Ling Chen, Ayano Satoh, David S. Salomon, Masaharu Seno

Biomaterials 301 122249 - 122249 2023年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

The heterogeneous cell population in the stromal microenvironment is considered to be attributed to the multiple sources from which the cells originate. Tumor associated myoepithelial cells (TAMEs) are one of the most important populations in the tumor microenvironment (TME) especially in breast cancer. On the other hand, cancer stem cells (CSCs) have previously been described to be the origin of tumor-associated cellular components in the TME. We prepared a cancer stem cell model converting mouse-induced pluripotent stem cells (miPSCs) in the presence of conditioned medium of breast cancer cell line MDA-MB-231 cells. The converted cells developed tumors progressing into invasive carcinoma with ductal carcinoma in situ (DCIS) like structure when transplanted into mouse mammary fat pads. The primary cultured cells from the tumor further exhibited markers of CSC such as Sox2, Oct3/4, - CD133 and EpCAM, and mammary gland-related TAME markers such as α-smooth muscle actin, cytokeratin 8, whey acidic protein, prolactin receptor and progesterone receptor as well. These results indicated that the CSCs could be an origin of TAMEs contributing to mammary gland epithelial cell differentiation and the progression to invasive carcinoma during tumor development. The gene expression profiles confirmed the enhanced signaling pathways of PI3K/AKT and MAPK, which have been demonstrated to be enriched in the CSC models, together with the estrogen receptor signaling which was peculiar to mammary gland-derived character.

DOI： 10.1016/j.biomaterials.2023.122249

The microbiota catabolites of quercetin glycosides concertedly enhance the resistance against acetaldehyde-induced oxidative stress 査読国際誌

Kexin Li, Hongyan Wu, Minori Kidawara, Yun Lin, Ayano Satoh, Gongliang Zhang, Shintaro Munemasa, Yoshiyuki Murata, Toshiyuki Nakamura, Yoshimasa Nakamura

Free Radical Research 56 ( 9-10 ) 607 - 616 2022年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Informa UK Limited

3,4-Dihydroxyphenylacetic acid (DOPAC) and 3-hydroxyphenylacetic acid (OPAC) are the predominant catabolites of quercetin glycosides, such as quercetin 4'-O-β-glucoside from the onion, produced by intestinal microbiota. Although each catabolite has been reported to protect the cells from acetaldehyde-induced cytotoxicity, the effect of their combination remains to be clarified. The purpose of this study was to determine whether the combination of DOPAC and OPAC enhances the resistance against the acetaldehyde-induced oxidative stress in the cultured hepatocytes. The pretreatment of the combination of DOPAC (5 μM) and OPAC (5 μM) showed significant protection against the acetaldehyde- and hydrogen peroxide-induced cytotoxicity, even though each compound at the same concentration did not. This combination also significantly inhibited the intracellular dichlorofluorescin diacetate-detectable reactive oxygen species (ROS) level, whereas the solo treatment did slightly, suggesting that reducing mechanisms of ROS or compounds that enhance ROS production are involved in the cytoprotective effect. The combinatory treatment significantly enhanced the gene expression of not only the aldehyde dehydrogenases (ALDHs), but also glutamate-cysteine ligase, catalytic subunit, the first rate-limiting enzyme of glutathione (GSH) synthesis. Accordingly, both the intracellular GSH level and the total ALDH activity were enhanced by DOPAC plus OPAC. Involvement of GSH in the cytoprotection as well as ALDH up-regulation by the combination was confirmed by the experiments using a GSH biosynthesis inhibitor, buthionine sulfoximine. Taken together, the present results suggested that the quercetin microbiota catabolites concertedly protect the cells from acetaldehyde through a pre-enhanced resistance against oxidative stress by the GSH-dependent up-regulation of ALDHs.

DOI： 10.1080/10715762.2022.2159820

Immune State Conversion of the Mesenteric Lymph Node in a Mouse Breast Cancer Model 査読国際誌

Tsukasa Shigehiro, Maho Ueno, Mayumi Kijihira, Ryotaro Takahashi, Chiho Umemura, Eman A. Taha, Chisaki Kurosaka, Megumi Asayama, Hiroshi Murakami, Ayano Satoh, Yoshimasa Nakamura, Junichiro Futami, Junko Masuda

International Journal of Molecular Sciences 23 ( 19 ) 11035 - 11035 2022年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

Secondary lymphoid tissues, such as the spleen and lymph nodes (LNs), contribute to breast cancer development and metastasis in both anti- and pro-tumoral directions. Although secondary lymphoid tissues have been extensively studied, very little is known about the immune conversion in mesenteric LNs (mLNs) during breast cancer development. Here, we demonstrate inflammatory immune conversion of mLNs in a metastatic 4T1 breast cancer model. Splenic T cells were significantly decreased and continuously suppressed IFN-γ production during tumor development, while myeloid-derived suppressor cells (MDSCs) were dramatically enriched. However, T cell numbers in the mLN did not decrease, and the MDSCs only moderately increased. T cells in the mLN exhibited conversion from a pro-inflammatory state with high IFN-γ expression to an anti-inflammatory state with high expression of IL-4 and IL-10 in early- to late-stages of breast cancer development. Interestingly, increased migration of CD103+CD11b+ dendritic cells (DCs) into the mLN, along with increased (1→3)-β-D-glucan levels in serum, was observed even in late-stage breast cancer. This suggests that CD103+CD11b+ DCs could prime cancer-reactive T cells. Together, the data indicate that the mLN is an important lymphoid tissue contributing to breast cancer development.

DOI： 10.3390/ijms231911035

Design and Synthesis of Glycosylated Cholera Toxin B Subunit as a Tracer of Glycoprotein Trafficking in Organelles of Living Cells 査読国際誌

Yuta Maki, Kazuki Kawata, Yanbo Liu, Kang‐Ying Goo, Ryo Okamoto, Yasuhiro Kajihara, Ayano Satoh

Chemistry – A European Journal 28 ( 37 ) e202201253 2022年5月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

Abstract

Glycosylation of proteins is known to be essential for changing biological activity and stability of glycoproteins on the cell surfaces and in body fluids. Delivering of homogeneous glycoproteins into the endoplasmic reticulum (ER) and the Golgi apparatus would enable us to investigate the function of asparagine‐linked (N‐) glycans in the organelles. In this work, we designed and synthesized an intentionally glycosylated cholera toxin B‐subunit (CTB) to be transported to the organelles of mammalian cells. The heptasaccharide, the intermediate structure of various complex‐type N‐glycans, was introduced to the CTB. The synthesized monomeric glycosyl‐CTB successfully entered mammalian cells and was transported to the Golgi and the ER, suggesting the potential use of synthetic CTB to deliver and investigate the functions of homogeneous N‐glycans in specific organelles of living cells.

DOI： 10.1002/chem.202201253

その他リンク： https://onlinelibrary.wiley.com/doi/full-xml/10.1002/chem.202201253

A Major Intestinal Catabolite of Quercetin Glycosides, 3-Hydroxyphenylacetic Acid, Protects the Hepatocytes from the Acetaldehyde-Induced Cytotoxicity through the Enhancement of the Total Aldehyde Dehydrogenase Activity 査読国際誌

Yujia Liu, Takumi Myojin, Kexin Li, Ayuki Kurita, Masayuki Seto, Ayano Motoyama, Xiaoyang Liu, Ayano SATOH, Shintaro Munemasa, Yoshiyuki Murata, Toshiyuki Nakamura, Yoshimasa Nakamura

International Journal of Molecular Sciences 23 ( 3 ) 1762 - 1762 2022年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：{MDPI} {AG}

Aldehyde dehydrogenases (ALDHs) are the major enzyme superfamily for the aldehyde metabolism. Since the ALDH polymorphism leads to the accumulation of acetaldehyde, we considered that the enhancement of the liver ALDH activity by certain food ingredients could help prevent alcohol-induced chronic diseases. Here, we evaluated the modulating effects of 3-hydroxyphenylacetic acid (OPAC), the major metabolite of quercetin glycosides, on the ALDH activity and acetaldehyde-induced cytotoxicity in the cultured cell models. OPAC significantly enhanced the total ALDH activity not only in mouse hepatoma Hepa1c1c7 cells, but also in human hepatoma HepG2 cells. OPAC significantly increased not only the nuclear level of aryl hydrocarbon receptor (AhR), but also the AhR-dependent reporter gene expression, though not the nuclear factor erythroid-2-related factor 2 (Nrf2)-dependent one. The pretreatment of OPAC at the concentration required for the ALDH upregulation completely inhibited the acetaldehyde-induced cytotoxicity. Silencing AhR impaired the resistant effect of OPAC against acetaldehyde. These results strongly suggested that OPAC protects the cells from the acetaldehyde-induced cytotoxicity, mainly through the AhR-dependent and Nrf2-independent enhancement of the total ALDH activity. Our findings suggest that OPAC has a protective potential in hepatocyte models and could offer a new preventive possibility of quercetin glycosides for targeting alcohol-induced chronic diseases.

DOI： 10.3390/ijms23031762

Identification of uncharacterized proteins potentially localized to mitochondria (UPMs) in <scp>Saccharomyces cerevisiae</scp> using a fluorescent protein unstable in the cytoplasm 査読国際誌

Satoshi Horiuchi, Shotaro Namba, Nozomu Saeki, Ayano Satoh, Hisao Moriya

Yeast 39 ( 5 ) 303 - 311 2021年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

Abstract

Eukaryotic cells are composed of organelles, and each organelle contains proteins that play a role in its function. Therefore, the localization of a protein, especially to organelles, is a clue to infer the function of that protein. In this study, we attempted to identify novel mitochondrially localized proteins in the budding yeast Saccharomyces cerevisiae using a fluorescent protein (GFPdeg) that is rapidly degraded in the cytoplasm. Of the budding yeast proteins predicted to localize to mitochondria by the prediction tool Deeploc‐1.0, those with known mitochondrial localization or functional relevance were eliminated, and 95 proteins of unknown function were selected as candidates for analysis. By forced expression of GFPdeg fusion proteins with these proteins and observation of their localization, we identified 35 uncharacterized proteins potentially localized to mitochondria (UPMs) including 8 previously identified proteins that localize to mitochondria. Most of these had no N‐terminal mitochondrial localization signal and were evolutionarily young “emerging genes” that exist only in S. cerevisiae. Some of these genes were found to be upregulated during the postdiauxic shift phase when mitochondria are being developed, suggesting that they are actually involved in some mitochondrial function.

DOI： 10.1002/yea.3685

その他リンク： https://onlinelibrary.wiley.com/doi/full-xml/10.1002/yea.3685

Enhanced cellular engraftment of adipose-derived mesenchymal stem cell spheroids by using nanosheets as scaffolds 査読国際誌

Hisato Nagano, Yoshitaka Suematsu, Megumi Takuma, Shimpo Aoki, Ayano Satoh, Eiji Takayama, Manabu Kinoshita, Yuji Morimoto, Shinji Takeoka, Toshinori Fujie, Tomoharu Kiyosawa

Scientific Reports 11 ( 1 ) 14500 - 14500 2021年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer Science and Business Media LLC

Abstract

The short survival time of transplanted adipose-derived mesenchymal stem cells (ASCs) is a problem for skin wound healing. Transplantation after the formation of cellular spheroids has been investigated as a promising method for prolonging cellular survival. However, there have been technical restrictions for transplantation of spheroids in clinical practice. Here, we show an effective method for transplantation of ASC spheroids onto skin wounds in order to efficiently cure refractory ulcers. To assist anchoring of spheroids onto skin wounds, we used a 120-nm-thick free-standing film (nanosheet) that has a highly adhesive property. Bioluminescence imaging showed that ASC spheroids carried by the nanosheet survived for 14 days, which is about two-times longer than that previously reported. Wounds treated with a nanosheet carrying ASC spheroids were 4-times smaller than untreated wounds on day 14. This method for transplantation of spheroids could be applied to cell therapy for various refractory skin wounds.

DOI： 10.1038/s41598-021-93642-6

その他リンク： https://www.nature.com/articles/s41598-021-93642-6

Horseradish peroxidase interacts with the cell wall peptidoglycans on oral bacteria 査読国際誌

Hirofumi Mizuno, Eiji Takayama, Ayano Satoh, Takeshi Into, Masanori Adachi, Daisuke Ekuni, Koji Yashiro, Masako Mizuno‑Kamiya, Motohiko Nagayama, Seitaro Saku, Takaaki Tomofuji, Yutaka Doi, Yukitaka Murakami, Nobuo Kondoh, Manabu Morita

Experimental and Therapeutic Medicine 20 ( 3 ) 2822 - 2827 2020年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Spandidos Publications

Salivary peroxidase and myeloperoxidase are known to display antibacterial activity against oral microbes, and previous indications have pointed to the possibility that horseradish peroxidase (HRP) adsorbs onto the membrane of the major oral streptococci, Streptococcus mutans and Streptococcus sanguinis (S. sanguinis). However, the mechanism of interaction between HRP and the bacterial cell wall component is unclear. Dental plaques containing salivary glycoproteins and extracellular microbial products are visualized with 'dental plaque disclosing agent', and are controlled within dental therapy. However, current 'dental plaque disclosing agents' are difficult to evaluate with just dental plaques, since they stain and disclose not only dental plaques but also pellicle formed with salivary glycoproteins on a tooth surface. In this present study, we have demonstrated that HRP interacted with the cell wall component of the major gram-positive bacterial peptidoglycan, but not the major cell wall component of gram-negative bacteria lipopolysaccharide. Furthermore, we observed that the adsorbed HRP labeled with fluorescence was detected on the major oral gram-positive strains S. sanguinis and Streptococcus salivarius (S. salivarius), but not on a gram-negative strain, Escherichia coli (E. coli). Furthermore, we have demonstrated that the combination of HRP and chromogenic substrate clearly disclosed the dental plaques and the biofilm developed by S. sanguinis, S. salivarius and the major gram-postive bacteria Lactobacillus casei on tooth surfaces, and slightly disclosed the biofilm by E. coli. The combination of HRP and chromogenic substrate did not stain either the dental pellicle with the salivary glycoprotein mucin, or naked tooth surfaces. These results have suggested the possibility that the adsorption activity of HRP not only contributes to the evaluation of dental plaque, but that enzymatic activity of HRP may also contribute to improve dental hygiene.

DOI： 10.3892/etm.2020.9016

Highly reliable, targeted photothermal cancer therapy combined with thermal dosimetry using a near-infrared absorbent 査読国際誌

Shinsuke Nomura, Yuji Morimoto, Hironori Tsujimoto, Masashi Arake, Manabu Harada, Daizoh Saitoh, Isao Hara, Eiichi Ozeki, Ayano Satoh, Eiji Takayama, Kazuo Hase, Yoji Kishi, Hideki Ueno

Scientific Reports 10 ( 1 ) 9765 - 9765 2020年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer Science and Business Media LLC

Abstract

Photothermal therapy (PTT) using a photo-absorbent in the near-infrared (NIR) region is an effective methodology for local cancer treatment. Before PTT using a NIR absorbent is executed, the operator generally determines the two parameters of fluence rate and irradiation time. However, even if the irradiation parameters are unchanged, the therapeutic effect of PTT is often different for individual tumors. Hence, we examined the therapeutic effect of PTT using a NIR absorbent (ICG lactosome) while changing two parameters (fluence rate and irradiation time) in various combinations. As a result, there was no robust correlation between those parameters and the therapeutic effect. Compared to those parameters, we found that a more reliable determinant was maintenance of the tumor temperature above 43 °C during NIR irradiation. To reconfirm the significance of the determinant, we developed a new system that can regulate the temperature at the NIR irradiation site at a constant level. By using the new system, we verified the treatment outcomes for tumors in which the NIR absorbent had accumulated. All of the tumors that had been kept at 43 °C during NIR irradiation were cured, while none of the tumors that had been kept at a temperature below 41 °C were cured. In conclusion, PTT using a NIR absorbent with thermal dosimetry is a highly reliable treatment for cancer.

DOI： 10.1038/s41598-020-66646-x

その他リンク： https://www.nature.com/articles/s41598-020-66646-x

Knockout of MMP3 Weakens Solid Tumor Organoids and Cancer Extracellular Vesicles 査読国際誌

Eman Taha, Chiharu Sogawa, Yuka Okusha, Hotaka Kawai, May Oo, Abdellatif Elseoudi, Yanyin Lu, Hitoshi Nagatsuka, Satoshi Kubota, Ayano Satoh, Kuniaki Okamoto, Takanori Eguchi

Cancers 12 ( 5 ) 1260 - 1260 2020年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

The tumor organoid (tumoroid) model in three-dimensional (3D) culture systems has been developed to reflect more closely the in vivo tumors than 2D-cultured tumor cells. Notably, extracellular vesicles (EVs) are efficiently collectible from the culture supernatant of gel-free tumoroids. Matrix metalloproteinase (MMP) 3 is a multi-functional factor playing crucial roles in tumor progression. However, roles of MMP3 within tumor growth and EVs have not unveiled. Here, we investigated the protumorigenic roles of MMP3 on integrities of tumoroids and EVs. We generated MMP3-knockout (KO) cells using the CRISPR/Cas9 system from rapidly metastatic LuM1 tumor cells. Moreover, we established fluorescent cell lines with palmitoylation signal-fused fluorescent proteins (tdTomato and enhanced GFP). Then we confirmed the exchange of EVs between cellular populations and tumoroids. LuM1-tumoroids released large EVs (200–1000 nm) and small EVs (50–200 nm) while the knockout of MMP3 resulted in the additional release of broken EVs from tumoroids. The loss of MMP3 led to a significant reduction in tumoroid size and the development of the necrotic area within tumoroids. MMP3 and CD9 (a category-1 EV marker tetraspanin protein) were significantly down-regulated in MMP3-KO cells and their EV fraction. Moreover, CD63, another member of the tetraspanin family, was significantly reduced only in the EVs fractions of the MMP3-KO cells compared to their counterpart. These weakened phenotypes of MMP3-KO were markedly rescued by the addition of MMP3-rich EVs or conditioned medium (CM) collected from LuM1-tumoroids, which caused a dramatic rise in the expression of MMP3, CD9, and Ki-67 (a marker of proliferating cells) in the MMP3-null/CD9-low tumoroids. Notably, MMP3 enriched in tumoroids-derived EVs and CM deeply penetrated recipient MMP3-KO tumoroids, resulting in a remarkable enlargement of solid tumoroids, while MMP3-null EVs did not. These data demonstrate that EVs can mediate molecular transfer of MMP3, resulting in increasing the proliferation and tumorigenesis, indicating crucial roles of MMP3 in tumor progression.

DOI： 10.3390/cancers12051260

Development of an experimental method of systematically estimating protein expression limits in HEK293 cells 査読国際誌

Yoshihiro Mori, Yuki Yoshida, Ayano Satoh, Hisao Moriya

Scientific Reports 10 ( 1 ) 4798 - 4798 2020年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer Science and Business Media LLC

Abstract

Protein overexpression sometimes causes cellular defects, although the underlying mechanism is still unknown. A protein’s expression limit, which triggers cellular defects, is a useful indication of the underlying mechanism. In this study, we developed an experimental method of estimating the expression limits of target proteins in the human embryonic kidney cell line HEK293 by measuring the proteins’ expression levels in cells that survived after the high-copy introduction of plasmid DNA by which the proteins were expressed under a strong cytomegalovirus promoter. The expression limits of nonfluorescent target proteins were indirectly estimated by measuring the levels of green fluorescent protein (GFP) connected to the target proteins with the self-cleaving sequence P2A. The expression limit of a model GFP was ~5.0% of the total protein, and sustained GFP overexpression caused cell death. The expression limits of GFPs with mitochondria-targeting signals and endoplasmic reticulum localization signals were 1.6% and 0.38%, respectively. The expression limits of four proteins involved in vesicular trafficking were far lower compared to a red fluorescent protein. The protein expression limit estimation method developed will be valuable for defining toxic proteins and consequences of protein overexpression.

DOI： 10.1038/s41598-020-61646-3

その他リンク： https://www.nature.com/articles/s41598-020-61646-3

Hematopoietic Cells Derived from Cancer Stem Cells Generated from Mouse Induced Pluripotent Stem Cells 査読国際誌

Ghmkin Hassan, SAID M AFIFY, Neha Nair, Kazuki Kumon, Amira Osman, Juan Du, Hager Mansour, Hagar A Abu Quora, Hend M Nawara, Ayano SATOH, Maram H. Zahra, Nobuhiro Okada, Akimasa Seno, Masaharu Seno

Cancers 12 ( 1 ) 2019年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

Cancer stem cells (CSCs) represent the subpopulation of cancer cells with the ability to differentiate into other cell phenotypes and initiated tumorigenesis. Previously, we reported generating CSCs from mouse induced pluripotent stem cells (miPSCs). Here, we investigated the ability of the CSCs to differentiate into hematopoietic cells. First, the primary cells were isolated from malignant tumors that were formed by the CSCs. Non-adherent cells (NACs) that arose from adherent cells were collected and their viability, as well as the morphology and expression of hematopoietic cell markers, were analyzed. Moreover, NACs were injected into the tail vein of busulfan conditioned Balb/c nude mice. Finally, CSCs were induced to differentiate to macrophages while using IL3 and SCF. The round nucleated NACs were found to be viable, positive for hematopoietic lineage markers and CD34, and expressed hematopoietic markers, just like homing to the bone marrow. When NACs were injected into mice, Wright-Giemsa staining showed that the number of white blood cells got higher than those in the control mice after four weeks. CSCs also showed the ability to differentiate toward macrophages. CSCs were demonstrated to have the potential to provide progenies with hematopoietic markers, morphology, and homing ability to the bone marrow, which could give new insight into the tumor microenvironment according to the plasticity of CSCs.

DOI： 10.3390/cancers12010082

Yeast screening system reveals the inhibitory mechanism of cancer cell proliferation by benzyl isothiocyanate through down-regulation of Mis12 査読国際誌

Naomi Abe-Kanoh, Narumi Kunisue, Takumi Myojin, Ayako Chino, Shintaro Munemasa, Yoshiyuki Murata, Ayano Satoh, Hisao Moriya, Yoshimasa Nakamura

Scientific Reports 9 ( 1 ) 8866 - 8866 2019年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer Science and Business Media LLC

Abstract

Benzyl isothiocyanate (BITC) is a naturally-occurring isothiocyanate derived from cruciferous vegetables. BITC has been reported to inhibit the proliferation of various cancer cells, which is believed to be important for the inhibition of tumorigenesis. However, the detailed mechanisms of action remain unclear. In this study, we employed a budding yeast Saccharomyces cerevisiae as a model organism for screening. Twelve genes including MTW1 were identified as the overexpression suppressors for the antiproliferative effect of BITC using the genome-wide multi-copy plasmid collection for S. cerevisiae. Overexpression of the kinetochore protein Mtw1 counteracts the antiproliferative effect of BITC in yeast. The inhibitory effect of BITC on the proliferation of human colon cancer HCT-116 cells was consistently suppressed by the overexpression of Mis12, a human orthologue of Mtw1, and enhanced by the knockdown of Mis12. We also found that BITC increased the phosphorylated and ubiquitinated Mis12 level with consequent reduction of Mis12, suggesting that BITC degrades Mis12 through an ubiquitin-proteasome system. Furthermore, cell cycle analysis showed that the change in the Mis12 level affected the cell cycle distribution and the sensitivity to the BITC-induced apoptosis. These results provide evidence that BITC suppresses cell proliferation through the post-transcriptional regulation of the kinetochore protein Mis12.

DOI： 10.1038/s41598-019-45248-2

その他リンク： https://www.nature.com/articles/s41598-019-45248-2

Suppression effect on IFN-γ of adipose tissue-derived mesenchymal stem cells isolated from β2-microglobulin-deficient mice. 査読国際誌

Junko Masuda, Eiji Takayama, Tatsuo Ichinohe, Warren Strober, Masako Mizuno-Kamiya, Tomokatsu Ikawa, Atsushi Kitani, Harumi Kawaki, Ivan Fuss, Hiroshi Kawamoto, Akimasa Seno, Arun Vaidyanath, Naoki Umemura, Akifumi Mizutani, Tomonari Kasai, Yasuko Honjo, Ayano Satoh, Hiroshi Murakami, Yoshimoto Katsura, Nobuo Kondoh, Masaharu Seno

Experimental and therapeutic medicine 16 ( 5 ) 4277 - 4282 2018年11月

記述言語：英語掲載種別：研究論文（学術雑誌）

Administration of bone marrow-derived mesenchymal stem cells (MSCs) is a possible treatment for graft-versus-host disease (GVHD) following allogeneic hematopoietic stem cell transplantation and other inflammatory conditions. To address the mechanism of immunosuppression by MSCs, in particular those derived from adipose tissue (AMSCs), AMSCs were isolated from three different mouse strains, and the suppressive capacity of the AMSCs thus obtained to suppress interferon (IFN)-γ generation in mixed lymphocyte reaction cultures serving as an in vitro model of GVHD were assessed. It was revealed that the AMSCs had a potent capacity to suppress IFN-γ production regardless of their strain of origin and that such suppression was not associated with production of interleukin-10. In addition, the results demonstrated that β2-microglobulin (β2m)-deficient AMSCs from β2m-/- mice were also potent suppressor cells, verifying the fact that the mechanism underlying the suppression by AMSCs is independent of major histocompatibility complex (MHC) class I expression or MHC compatibility. As AMSCs appear to have immunosuppressive properties, AMSCs may be a useful source of biological suppressor cells for the control of GVHD in humans.

DOI： 10.3892/etm.2018.6689

Cytokine Expression and Macrophage Localization in Xenograft and Allograft Tumor Models Stimulated with Lipopolysaccharide. 査読国際誌

Junko Masuda, Tsukasa Shigehiro, Takuma Matsumoto, Ayano Satoh, Akifumi Mizutani, Chiho Umemura, Shoki Saito, Mayumi Kijihira, Eiji Takayama, Akimasa Seno, Hiroshi Murakami, Masaharu Seno

International journal of molecular sciences 19 ( 4 ) 2018年4月

記述言語：英語掲載種別：研究論文（学術雑誌）

T cell-deficient mice such as nude mice are often used to generate tumor xenograft for the development of anticancer agents. However, the functionality of the other immune cells including macrophages, dendritic cells (DCs), and myeloid-derived suppressor cells (MDSCs) in the xenograft are largely unknown. Macrophages and dendritic cells (DCs) acquire functionally distinct properties in response to various environmental stimuli; the interaction of these cells with MDSCs in tumor microenvironments regulates cancer progression. Nude mice are less likely to reject human cancer cells because of major histocompatibility complex (MHC) mismatches. The tumor microenvironment in a xenograft, comprising human and mouse cells, exhibits more complex bidirectional signaling and function than that of allograft. Here, we evaluated the differences of myeloid cells between them. Plasma interferon-γ and interleukin-18 concentrations in the xenograft tumor model after lipopolysaccharide (LPS) administration were significantly higher than those in the allograft tumor model. MHC class I, II, and CD80 expression levels were increased in CD11b⁺ and MDSC populations after LPS administration in the spleen of a xenograft tumor model but not in that of an allograft tumor model. Additionally, the number of CD80- and mannose receptor C type 1 (MRC1)-expressing cells was decreased upon LPS administration in the tumor of the xenograft tumor. These results suggest that functions of macrophages and DCs are sustained in the xenograft, whereas their functions in response to LPS were suppressed in the allograft. The findings will encourage the consideration of the effects of myeloid cells in the xenograft for drug development.

DOI： 10.3390/ijms19041261

Practical Liposomal Formulation for Taxanes with Polyethoxylated Castor Oil and Ethanol with Complete Encapsulation Efficiency and High Loading Efficiency 査読国際誌

Tsukasa Shigehiro, Junko Masuda, Shoki Saito, Apriliana Khayrani, Kazumasa Jinno, Akimasa Seno, Arun Vaidyanath, Akifumi Mizutani, Tomonari Kasai, Hiroshi Murakami, Ayano Satoh, Tetsuya Ito, Hiroki Hamada, Yuhki Seno, Tadakatsu Mandai, Masaharu Seno

Nanomaterials 7 ( 10 ) 290 - 290 2017年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

Taxanes including paclitaxel and docetaxel are effective anticancer agents preferably sufficient for liposomal drug delivery. However, the encapsulation of these drugs with effective amounts into conventional liposomes is difficult due to their high hydrophobicity. Therefore, an effective encapsulation strategy for liposomal taxanes has been eagerly anticipated. In this study, the mixture of polyethoxylated castor oil (Cremophor EL) and ethanol containing phosphate buffered saline termed as CEP was employed as a solvent of the inner hydrophilic core of liposomes where taxanes should be incorporated. Docetaxel-, paclitaxel-, or 7-oxacetylglycosylated paclitaxel-encapsulating liposomes were successfully prepared with almost 100% of encapsulation efficiency and 29.9, 15.4, or 29.1 mol% of loading efficiency, respectively. We then applied the docetaxel-encapsulating liposomes for targeted drug delivery. Docetaxel-encapsulating liposomes were successfully developed HER2-targeted drug delivery by coupling HER2-specific binding peptide on liposome surface. The HER2-targeting liposomes exhibited HER2-specific internalization and enhanced anticancer activity in vitro. Therefore, we propose the sophisticated preparation of liposomal taxanes using CEP as a promising formulation for effective cancer therapies.

DOI： 10.3390/nano7100290

Tumor growth limited to subcutaneous site vs tumor growth in pulmonary site exhibit differential effects on systemic immunities 査読国際誌

Junko Masuda, Eiji Takayama, Warren Strober, Ayano Satoh, Yuji Morimoto, Yasuko Honjo, Tatsuo Ichinohe, Shin-Ichi Tokuno, Toshiaki Ishizuka, Takahiro Nakata, Akifumi Mizutani, Naoki Umemura, Atsushi Kitani, Ivan J. Fuss, Tsukasa Shigehiro, Harumi Kawaki, Masako Mizuno-Kamiya, Nobuo Kondoh, Masaharu Seno

Oncology Reports 38 ( 1 ) 449 - 455 2017年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Spandidos Publications

To evaluate systemic immunity associated with tumor growth limited to a subcutaneous site versus growth proceeding at multiple tumor sites, we established syngeneic mouse subcutaneous and pulmonary tumor models by local subcutaneous and intravenous injection of colon carcinoma CT26 cells. We found that splenic myeloid-derived suppressor cell (MDSC) levels were significantly increased in the subcutaneous tumor model but not in the pulmonary tumor model. Furthermore, both CD4+ and CD8+ T cells as well as CD4+ Foxp3+ T cells were significantly decreased in the subcutaneous tumor model and were largely unchanged in the pulmonary tumor model. In addition, the subcutaneous model, but not the pulmonary model, displayed a Th1 polarization bias. This bias was characterized by decreased IL-4, IL-9, and IL-10 production, whereas the pulmonary model displayed increased production of IL-10. These results suggest that the mode of tumor development has differential effects on systemic immunity that may, in turn, influence approaches to treatment of cancer patients.

DOI： 10.3892/or.2017.5646

Do the Golgi Glycosylation Enzymes Cycle between the Endoplasmic Reticulum and the Golgi Apparatus? 査読

Ayano Satoh, Yasuko Honjo

Trends in Glycoscience and Glycotechnology 29 ( 167 ) E49 - E50 2017年

Transient Tcf3 Gene Repression by TALE-Transcription Factor Targeting 査読国際誌

Junko Masuda, Hiroshi Kawamoto, Warren Strober, Eiji Takayama, Akifumi Mizutani, Hiroshi Murakami, Tomokatsu Ikawa, Atsushi Kitani, Narumi Maeno, Tsukasa Shigehiro, Ayano Satoh, Akimasa Seno, Vaidyanath Arun, Tomonari Kasai, Ivan J. Fuss, Yoshimoto Katsura, Masaharu Seno

Applied Biochemistry and Biotechnology 180 ( 8 ) 1559 - 1573 2016年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer Science and Business Media LLC

Transplantation of hematopoietic stem and progenitor cells (HSCs) i.e., self-renewing cells that retain multipotentiality, is now a widely performed therapy for many hematopoietic diseases. However, these cells are present in low number and are subject to replicative senescence after extraction; thus, the acquisition of sufficient numbers of cells for transplantation requires donors able to provide repetitive blood samples and/or methods of expanding cell numbers without disturbing cell multipotentiality. Previous studies have shown that HSCs maintain their multipotentiality and self-renewal activity if TCF3 transcription function is blocked under B cell differentiating conditions. Taking advantage of this finding to devise a new approach to HSC expansion in vitro, we constructed an episomal expression vector that specifically targets and transiently represses the TCF3 gene. This consisted of a vector encoding a transcription activator-like effector (TALE) fused to a Krüppel-associated box (KRAB) repressor. We showed that this TALE-KRAB vector repressed expression of an exogenous reporter gene in HEK293 and COS-7 cell lines and, more importantly, efficiently repressed endogenous TCF3 in a human B lymphoma cell line. These findings suggest that this vector can be used to maintain multipotentiality in HSC being subjected to a long-term expansion regimen prior to transplantation.

DOI： 10.1007/s12010-016-2187-4

その他リンク： http://link.springer.com/article/10.1007/s12010-016-2187-4/fulltext.html

Transport within the Golgi: For the Study of Glycoprotein Movement 2 査読

Ayano Satoh, Yasuko Honjo

Trends in Glycoscience and Glycotechnology 28 ( 161 ) E61 - E62 2016年

Comparative Analysis of Cartilage Marker Gene Expression Patterns during Axolotl and Xenopus Limb Regeneration 査読国際誌

Kazumasa Mitogawa, Aki Makanae, Ayano Satoh, Akira Satoh

PLOS ONE 10 ( 7 ) e0133375 - e0133375 2015年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Public Library of Science (PLoS)

Axolotls (Ambystoma mexicanum) can completely regenerate lost limbs, whereas Xenopus laevis frogs cannot. During limb regeneration, a blastema is first formed at the amputation plane. It is thought that this regeneration blastema forms a limb by mechanisms similar to those of a developing embryonic limb bud. Furthermore, Xenopus laevis frogs can form a blastema after amputation; however, the blastema results in a terminal cone-shaped cartilaginous structure called a "spike." The causes of this patterning defect in Xenopus frog limb regeneration were explored. We hypothesized that differences in chondrogenesis may underlie the patterning defect. Thus, we focused on chondrogenesis. Chondrogenesis marker genes, type I and type II collagen, were compared in regenerative and nonregenerative environments. There were marked differences between axolotls and Xenopus in the expression pattern of these chondrogenesis-associated genes. The relative deficit in the chondrogenic capacity of Xenopus blastema cells may account for the absence of total limb regenerative capacity.

DOI： 10.1371/journal.pone.0133375

The Roles of GRASP55/65 in Golgi Formation and Function 査読

Ayano Satoh, Youko Hasegawa, Yasuko Honjo

Trends in Glycoscience and Glycotechnology 27 ( 153 ) 1 2015年1月

Cancer stem cells maintain a hierarchy of differentiation by creating their niche 査読国際誌

Shuichi Matsuda, Ting Yan, Akifumi Mizutani, Tatsuyuki Sota, Yuki Hiramoto, Marta Prieto‐Vila, Ling Chen, Ayano Satoh, Takayuki Kudoh, Tomonari Kasai, Hiroshi Murakami, Li Fu, David S. Salomon, Masaharu Seno

International Journal of Cancer 135 ( 1 ) 27 - 36 2013年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

The self‐renewal and differentiation properties of cancer stem cells (CSCs) are regulated and maintained by the CSC niche. However, the mechanism of this maintenance, especially the maintenance contributed by differentiated cancer cells, remains to be fully elucidated. Recently, we have established a model of CSCs, miPS‐LLCcm, from mouse induced pluripotent stem cells (miPSCs). In vitro cultured miPS‐LLCcm cells were autonomously balanced with stem‐like cells and differentiated cells including vascular endothelial cells. Under these conditions, the CSC properties appeared to be stable in the presence of the factor(s) secreted by the differentiated cells. The factor(s) activated Notch signaling and promoted self‐renewal of CSCs. In addition, the secreted factor(s) appeared to regulate the differentiation lineage of CSCs. Our results indicate that the differentiated progenies of CSCs containing vascular endothelium play important roles for regulating the CSC's properties. Therefore, miPS‐LLCcm cells create their own in vitro niche to maintain themselves in the hierarchy of differentiating CSCs.

DOI： 10.1002/ijc.28648

その他リンク： https://onlinelibrary.wiley.com/doi/full-xml/10.1002/ijc.28648

Identification of Caveolin-1 as a Potential Causative Factor in the Generation of Trastuzumab Resistance in Breast Cancer Cells 査読国際誌

Sreeja C Sekhar, Tomonari Kasai, Ayano Satoh, Tsukasa Shigehiro, Akifumi Mizutani, Hiroshi Murakami, Bishoy YA El-Aarag, Salomon DS, Anna Massaguer, Rafael de Llorens, Masaharu Seno

Journal of Cancer 4 ( 5 ) 391 - 401 2013年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Ivyspring International Publisher

The oncogenic tyrosine kinase receptor ErbB2 is a prognostic factor and target for breast cancer therapeutics. In contrast with the other ErbB receptors, ErbB2 is hardly internalized by ligand induced mechanisms, indicating a prevalent surface expression. Elevated levels of ErbB2 in tumor cells are associated with its defective endocytosis and down regulation. Here we show that caveolin-1 expression in breast cancer derived SKBR-3 cells (SKBR-3/Cav-1) facilitates ligand induced ErbB2 endocytosis using an artificial peptide ligand EC-eGFP. Similarly, stimulation with humanized anti ErbB2 antibody Trastuzumab (Herceptin) was found to be internalized and co-localized with caveolin-1 in SKBR-3/Cav-1 cells. Internalized EC-eGFP and Trastuzumab in SKBR-3/Cav-1 cells were then delivered via caveolae to the caveolin-1 containing early endosomes. Consequently, attenuated Fc receptor mediated ADCC functions were observed when exposed to Trastuzumab and EC-Fc (EC-1 peptide conjugated to Fc part of human IgG). On the other hand, this caveolae dependent endocytic synergy was not observed in parental SKBR-3 cells. Therefore, caveolin-1 expression in breast cancer cells could be a predictive factor to estimate how cancer cells are likely to respond to Trastuzumab treatment.

DOI： 10.7150/jca.6470

A Model of Cancer Stem Cells Derived from Mouse Induced Pluripotent Stem Cells 査読国際誌

Ling Chen, Tomonari Kasai, Yueguang Li, Yuh Sugii, Guoliang Jin, Masashi Okada, Arun Vaidyanath, Akifumi Mizutani, Ayano Satoh, Takayuki Kudoh, Mary J. C. Hendrix, David S. Salomon, Li Fu, Masaharu Seno

PLoS ONE 7 ( 4 ) e33544 - e33544 2012年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Public Library of Science (PLoS)

Cancer stem cells (CSCs) are capable of continuous proliferation and self-renewal and are proposed to play significant roles in oncogenesis, tumor growth, metastasis and cancer recurrence. CSCs are considered derived from normal stem cells affected by the tumor microenvironment although the mechanism of development is not clear yet. In 2007, Yamanaka's group succeeded in generating Nanog mouse induced pluripotent stem (miPS) cells, in which green fluorescent protein (GFP) has been inserted into the 5'-untranslated region of the Nanog gene. Usually, iPS cells, just like embryonic stem cells, are considered to be induced into progenitor cells, which differentiate into various normal phenotypes depending on the normal niche. We hypothesized that CSCs could be derived from Nanog miPS cells in the conditioned culture medium of cancer cell lines, which is a mimic of carcinoma microenvironment. As a result, the Nanog miPS cells treated with the conditioned medium of mouse Lewis lung carcinoma acquired characteristics of CSCs, in that they formed spheroids expressing GFP in suspension culture, and had a high tumorigenicity in Balb/c nude mice exhibiting angiogenesis in vivo. In addition, these iPS-derived CSCs had a capacity of self-renewal and expressed the marker genes, Nanog, Rex1, Eras, Esg1 and Cripto, associated with stem cell properties and an undifferentiated state. Thus we concluded that a model of CSCs was originally developed from miPS cells and proposed the conditioned culture medium of cancer cell lines might perform as niche for producing CSCs. The model of CSCs and the procedure of their establishment will help study the genetic alterations and the secreted factors in the tumor microenvironment which convert miPS cells to CSCs. Furthermore, the identification of potentially bona fide markers of CSCs, which will help the development of novel anti-cancer therapies, might be possible though the CSC model.

DOI： 10.1371/journal.pone.0033544

Enhanced internalization of ErbB2 in SK-BR-3 cells with multivalent forms of an artificial ligand 査読国際誌

Arun Vaidyanath, Toshihiro Hashizume, Tadahiro Nagaoka, Nao Takeyasu, Hitomi Satoh, Ling Chen, Jiyou Wang, Tomonari Kasai, Takayuki Kudoh, Ayano Satoh, Li Fu, Masaharu Seno

Journal of Cellular and Molecular Medicine 15 ( 11 ) 2525 - 2538 2011年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

Targeting and down-regulation of ErbB2, a member of EGF receptor family, is regarded as one of the key aspect for cancer treatment because it is often overexpressed in breast and ovarian cancer cells. Although natural ligands for ErbB2 have not been found, unlike other ErbB receptors, EC-1, a 20-amino acid circular peptide, has been shown to bind to ErbB2 as an artificial ligand. Previously we showed EC-1 peptide did not induce the internalization of ErbB2 in SK-BR-3 cells. In this report, we designed divalent and multivalent forms of EC-1 peptide with the Fc portion of the human IgG and bionanocapsule modified with ZZ-tag on its surface to improve the interaction with ErbB2. These forms showed higher affinity to ErbB2 than that of EC-1 monomer. Furthermore, prominent endosomal accumulation of ErbB2 occurred in SK-BR-3 cells when stimulated with EC-Fc ligand multivalently displayed on the surface of the bionanocapsule, whereas SK-BR-3 cells as themselves displayed stringent mechanism against ErbB2 internalization without stimulation. The multivalent form of EC-1 peptide appeared to internalize ErbB2 more efficiently than divalent form did. This internalization was unaffected by the inhibition of clathrin association, but inhibited when the cholesterol was depleted which explained either caveolar or GPI-AP-early endocytic compartment (GEEC) pathway. Because of the lack of caveolin-1 expression, caveolar machinery may be lost in SK-BR-3 cell line. Therefore, it is suggested that the multivalent form of EC-1 induces the internalization of ErbB2 through the GEEC pathway.

DOI： 10.1111/j.1582-4934.2011.01277.x

Beta-2-glycoprotein I and urinary trypsin inhibitor levels in the plasma of pregnant and postpartum women 査読国際誌

Junko Masuda, Kimihiro Suzuki, Ayano Satoh, Kyoko Kojima-Aikawa, Kuniaki Nakanishi, Koichi Kuroda, Mitsutaka Murakami, Eiji Takayama, Isamu Matsumoto

Thrombosis Research 117 ( 3 ) 255 - 261 2006年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

Annexins (Anx) are a family of structurally related proteins that all bind to anionic phospholipids in a Ca(2+)-dependent manner. Some biological properties of beta-2-glycoprotein I (beta(2)-GPI) are similar to those of Anx IV and Anx V. Urinary trypsin inhibitor (UTI) helps to maintain normal pregnancy and prevent preterm delivery by inhibiting uterine contraction. However, plasma beta(2)-GPI and UTI levels have not been measured in normal pregnancy. The aim of this study is to clarify the levels of these parameters. Subjects were nonpregnant women (n=50), 120 pregnant women, and maternal subjects just after delivery (n=53) or postpartum (n=67). All of the subjects were healthy. Plasma levels of beta(2)-GPI, UTI, Anx IV, Anx V and other coagulation and fibrinolysis markers were measured by ELISA. The mean plasma level of beta(2)-GPI was significantly increased during the third trimester of pregnancy and 3 to 5 days after delivery. The mean plasma level of UTI was unchanged from the first trimester of pregnancy to the postpartum period. The mean plasma UTI level in vaginal delivery group was significantly higher than that in cesarean section group. beta(2)-GPI protein was expressed in some of the syncytiotrophoblasts. These data suggest that beta(2)-GPI might act to prevent blood clotting on the placental surfaces and also prevents disseminated intravascular coagulation in the microcirculation and maternal plasma. UTI levels might be kept constant by increased urinary excretion despite overproduction during pregnancy.

DOI： 10.1016/j.thromres.2005.03.026

Annexins I and IV inhibit Staphylococcus aureus attachment to human macrophages 査読国際誌

Mari Gotoh, Yukiko Takamoto, Kahori Kurosaka, Junko Masuda, Michiru Ida, Ayano Satoh, Eiji Takayama, Kyoko Kojima-Aikawa, Yoshiro Kobayashi, Isamu Matsumoto

Immunology Letters 98 ( 2 ) 297 - 302 2005年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

Annexins are a family of proteins that bind to phospholipids and carbohydrates in a calcium-dependent manner. They are present in a variety of body fluids. Previous studies have shown that annexins have anti-inflammatory activities for lipid A of Gram-negative bacteria. The present study investigated the effect of annexins on interaction between Gram-positive bacteria and immune cells such as macrophages. Annexins I and IV bound to lipoteichoic acids which are surface molecules on Gram-positive bacteria. Binding of annexins I and IV to whole Staphylococcus aureus (S. aureus) were observed and these bindings were inhibited by lipoteichoic acid from S. aureus. Moreover, annexins I and IV suppressed the attachment of S. aureus to phorbol 12-myristate 13-acetate-treated THP-1 cells (human macrophages). These results suggest that annexins I and IV have ligand specificities toward foreign substances, and that the annexins might have some anti-inflammatory property for Gram-positive bacteria.

DOI： 10.1016/j.imlet.2004.12.004

A novel expression vector, designated as pHisJM, for producing recombinant His-fusion proteins 査読国際誌

Junko Masuda, Eiji Takayama, Ayano Satoh, Kyoko Kojima-Aikawa, Kimihiro Suzuki, Isamu Matsumoto

Biotechnology Letters 26 ( 20 ) 1543 - 1548 2004年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer Science and Business Media LLC

Compared to glutathione S -transferase (GST), tagging with hexahistidine residues (His) has several merits: low levels of toxicity and immunogenicity, a smaller size and no electric charge. We have constructed a novel expression vector, designated as pHisJM (EMBL/GenBank/DDJB accession no. AB116367), for producing recombinant His-fusion proteins. This vector was constructed by replacing GST and multiple cloning site (MCS) cassettes in pGEX-5X-3 with those of hexahistidine and MCS derived from pRSET C vector. Human annexin IV (Anx IV) was used as target protein. His-Anx IV fusion protein was expressed using pHisJM and gave a 40 kDa band when immuno-stained with anti-His mAb or anti-Anx IV mAb as predicted. To compare expression efficiency, a Anx IV cDNA inserted-pHisJM or pGEX-5X-3 was transformed into Escherichia coli DH5alpha, JM109, BL21 and BL21(DE3). Using pHisJM, Anx IV protein was highly expressed in all cell strains. In addition to the merits of using His-tag, pHisJM has several advantages: 1) it has high expression efficiency; 2) it can be used in any Escherichia coli strain; and 3) it can be used in a single strain of Escherichia coli in all steps from plasmid construction to the expression of the target gene.

DOI： 10.1023/b:bile.0000045650.90384.b2

Human Annexin V Binds to Sulfatide: Contribution to Regulation of Blood Coagulation 査読国際誌

M. Ida

Journal of Biochemistry 135 ( 5 ) 583 - 588 2004年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Oxford University Press (OUP)

Annexin V is a calcium-dependent phospholipid-binding protein that exhibits anticoagulant activity on binding to phosphatidylserine exposed on the activated surfaces of endothelial cells and platelets, inhibiting activation of factor X and prothrombin in the blood coagulation cascade. Sulfatide (galactosylceramide I(3)-sulfate), one of the glycosphingolipids of the platelet cell membrane, is thought to be involved in blood coagulation systems via activation of factor XII. In this study, we examined whether or not annexin V binds to sulfatide and affects the coagulant activity of sulfatide. Solid phase assaying of annexin V revealed that it binds specifically to sulfatide, i.e. not to galactosylceramide or gangliosides, in the presence of calcium ions. Affinity analysis by means of surface plasmon resonance showed that the K(D) of the interaction between annexin V and sulfatide is 1.2 micro M. Kinetic turbidometric assaying of plasma coagulation initiated by CaCl(2) revealed that the coagulation rate in the presence of sulfatide or phosphatidylserine was decreased by annexin V. These results suggest that annexin V regulates coagulability in the blood stream by binding not only to phosphatidylserine but also to sulfatide.

DOI： 10.1093/jb/mvh071

Levels of annexin IV and V in the plasma of pregnant and postpartum women 査読国際誌

Junko Masuda, Eiji Takayama, Ayano Satoh, Michiru Ida, Tadashi Shinohara, Kyoko Kojima-Aikawa, Fumitaka Ohsuzu, Kuniaki Nakanishi, Koichi Kuroda, Mitsutaka Murakami, Isamu Matsumoto, Kimihiro Suzuki

Thrombosis and Haemostasis 91 ( 06 ) 1129 - 1136 2004年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Georg Thieme Verlag KG

Summary

Annexin (Anx) V is pivotal in the maintenance of pregnancy by preventing the activation of blood coagulation. The homology of the amino acid sequence between Anx IV and Anx V is highest in Anx family proteins. However, little is known about the roles of Anx IV in pregnancy.The aim of this study is to clarify the roles of circulating Anx IV and Anx V in normal pregnancy. Subjects were non-pregnant women (n = 50), 120 pregnant women, and maternal subjects just after delivery (n = 53) or postpartum (n = 67). Anx IV in the plasma of non-pregnant women was at a concentration 20 times that of Anx V. The plasma levels of Anx IV suddenly increase after delivery, but Anx V levels remain low during this period. Anx IV and Anx V exert similar levels of anticoagulant activity. Anx IV protein was expressed on the basal surface of syncytiotrophoblasts; Anx V protein, on the apical surface of syncytiotrophoblasts. These results suggest that Anx IV enters the maternal bloodstream just after delivery and might play a role in preventing disseminated intravascular coagulopathy, and that Anx V helps to prevent clotting in the placenta during pregnancy.

DOI： 10.1160/th03-12-0778

Ligand-Binding Properties of Annexin from Caenorhabditis elegans (Annexin XVI, Nex-1) 査読

A. Satoh, M. Hazuki, K. Kojima, J. Hirabayashi, I. Matsumoto

Journal of Biochemistry 128 ( 3 ) 377 - 381 2000年9月

New role of glycosaminoglycans on the plasma membrane proposed by their interaction with phosphatidylcholine 査読

Ayano Satoh, Toshihiko Toida, Keiichi Yoshida, Kyoko Kojima, Isamu Matsumoto

FEBS Letters 477 ( 3 ) 249 - 252 2000年7月

担当区分：筆頭著者掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley

Glycosaminoglycan side chains of membrane proteoglycans have been claimed to be located at the outermost layer of the glycocalyx surrounding the cell. In this study measurements by surface plasmon resonance and solid‐phase assay have shown that both chondroitin sulfate and keratan sulfate but not heparin associate with phosphatidylcholine under physiological conditions. Spectrophotometric measurements also showed that chondroitin sulfate restricts the lateral diffusion of phosphatidylcholine in liposomes. These findings indicate that chondroitin sulfate and/or keratan sulfate chains of membrane proteoglycans crouch on the surface of the membrane while heparan sulfate chains stretch outward from the membrane surface as postulated traditionally.

DOI： 10.1016/s0014-5793(00)01746-4

Adsorption of anti-annexin V using dextran sulfate bound cellulose beads 査読

Kimihiro Suzuki, Ayano Satoh, Toshihiko Hidaka, Eiji Takayama, Kouji Kataharada, Mitsuyo Matsumoto, Tadashi Shinohara, Isamu Matsumoto, Fumitaka Ohsuzu

Journal of Clinical Apheresis 15 ( 4 ) 262 - 265 2000年

Comparison of Methods of Immobilization to Enzyme-Linked Immunosorbent Assay Plates for the Detection of Sugar Chains 査読

Ayano Satoh, Emiko Fukui, Saori Yoshino, Mayumi Shinoda, Kyoko Kojima, Isamu Matsumoto

Analytical Biochemistry 275 ( 2 ) 231 - 235 1999年11月

Analysis of Interaction between Lectin and Carbohydrate by Surface Plasmon Resonance 査読

Ayano Satoh, Isamu Matsumoto

Analytical Biochemistry 275 ( 2 ) 268 - 270 1999年11月

Detection of anti-Annexin IV and V antibodies in patients with antiphospholipid syndrome and systemic lupus erythematosus 査読

A Satoh, K Suzuki, E Takayama, K Kojima, T Hidaka, M Kawakami, Matsumoto, I, F Ohsuzu

JOURNAL OF RHEUMATOLOGY 26 ( 8 ) 1715 - 1720 1999年8月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：J RHEUMATOL PUBL CO

Objective, Annexins (Anx) are a family of structurally related proteins that bind to phospholipids in a calcium dependent manner. It has been reported that antibodies to Anx V, which acts as an antithrombotic protein, are associated with thrombosis in systemic lupus erythematosus (SLE) and/or antiphospholipid syndrome (APS). Homology between the primary structures of Anx IV and Anx V is the highest among members of the Anx family. We investigated whether anti-Anx IV autoantibodies can be detected in the sera of patients with SLE and/or APS. Methods. Seventy-four patients with SLE/APS were divided into 3 groups: Group A, patients with SLE but no clinical or serological features of APS; Group B, patients with SLE having only serological signs of APS; and Group C, patients with clinical symptoms and serological signs of AP, AN;IV and Anx V were prepared by recombinant technique. Anti-Anx IV, Anx V, cardiolipin (CL), and CL beta(2)-glycoprotein I were detected by ELISA. Results. Anti-Anx IV was found in 15.4% of Group A, 20.0% of Group B, and 21.7% of Group C, Anti-Anx V was found in 3.8% of Group A, 28.0% of Group B, and 30.4% of Group C. Significant correlations were noted between anti-Anx IV titer and anti-Anx V titer (p < 0.001), and between anti-Anx IV titer and aCL titer (p < 0.01). Conclusion. Anti-Anx TV and V antibodies were characterized in the sera of patients with SLE/APS. Significantly higher frequency of arterial or venous thrombosis was found in patients with anti-Anx V.

Immobilization of Saccharides and Peptides on 96-Well Microtiter Plates Coated with Methyl Vinyl Ether–Maleic Anhydride Copolymer 査読

Ayano Satoh, Kyoko Kojima, Tamami Koyama, Haruko Ogawa, Isamu Matsumoto

Analytical Biochemistry 260 ( 1 ) 96 - 102 1998年6月

Modulation of cell surface lectin receptors on K562 human erythroleukemia cells induced by transfection with annexin IV cDNA 査読

Ayano Satoh, Eiji Takayama, Kyoko Kojima, Haruko Ogawa, Yoshimoto Katsura, Tatsuo Kina, Tatsuro Irimura, Isamu Matsumoto

FEBS Letters 405 ( 1 ) 107 - 110 1997年3月

Characterization of Human p33/41 (Annexin IV), a Ca2+ Dependent Carbohydrate-Binding Protein with Monoclonal Anti-annexin IV Antibodies, AS11 and AS17. 査読

Ayano SATOH, Eiji TAKAYAMA, Kyoko KOJIMA, Haruko OGAWA, Yoshimoto KATSURA, Tatsuo KINA, Isamu MATSUMOTO

Biological and Pharmaceutical Bulletin 20 ( 3 ) 224 - 229 1997年

Expression of Carbohydrate-Binding Protein p33/41 in Human Tumor Cell Lines 査読

A. Satoh, E. Takayama, K. Kojima, H. Ogawa, T. Yamori, S. Sato, T. Kawaguchi, T. Tsuruo, Y. Katsura, T. Kina, I. Matsumoto

Journal of Biochemistry 119 ( 2 ) 346 - 353 1996年2月

　前のページ - 次のページ　

1