研究者詳細 - 島内　寿徳

MISC - 島内　寿徳

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Detection of Protein Conformation under Stress Condition Using Liposome as Sensor Materials

2004年

メンブレン・ストレスバイオテクノロジー--薬物送達システムの設計と開発への応用 (特集大きな可能性を秘めた研究開発)

久保井亮一, 馬越大, 島内寿徳

ケミカルエンジニヤリング 49 ( 4 ) 255 - 260 2004年

メンブレン・ストレスバイオテクノロジー--薬物送達システムの設計と開発への応用 (特集大きな可能性を秘めた研究開発)

久保井亮一, 馬越大, 島内寿徳

ケミカルエンジニヤリング 49 ( 4 ) 255 - 260 2004年

Metal Affinity-Immobilized Liposome Chromatography for Stress Mediated Separation of Biomolecules

Nagami Hideto, Umakoshi Hiroshi, Kitaura Takeaki, Thompson III Gary Lee, Shimanouchi Toshinori, Kuboi Ryoichi

アジア・太平洋化学工学会議発表論文要旨集 2004 991 - 991 2004年

記述言語：英語出版者・発行元：公益社団法人化学工学会

Investigation and utilization of peptides and proteins depend upon their separation and purification, which may be accomplished with chromatographic methods. Among the possible methods, immobilized metal affinity chromatography (IMAC), first reported by Porath, is known as a useful and powerful tool of the separation and analysis of the peptides and proteins and currently becomes the most popular chromatographic technique. A metal-affinity immobilized liposome chromatography (MA-ILC) was newly developed as a new chromatographic technique to separate and analyze the protein. The MA-ILC was prepared from the immobilized liposome modified with the functional ligands. A N-hexadodecyl -imonodiacetic acid (HIDA) was then used as the functional ligand. The metal ion Cu(II) was first adsorbed by the coordination with the functional ligand-modified liposome, immobilized on a gel matrix before the analysis of the elution behaviors. Synthetic peptides ranging in size from 5 to 40 residues were used to evaluate the retention properties of MA-ILC. The retention properties using usual imidazole elution for MA-ILC at 303 K was compared with those for IMAC, which is a most popular separation method for proteins, and an immobilized liposome chromatography (ILC), which can separate and analyze the protein mainly by the hydrophobic interaction between the liposome and protein. The retention ability of peptide on MA-ILC has both the properties of IMAC and ILC; the capacity factor depends on both the number of His residue in peptide and the hydrophobicity of peptide. It is considered that the retention properties of IMAC could be related not only to the hydrophobic interaction between the peptide and liposome but also to the cluster formation of the ligands on the liposome surface.

DOI： 10.11491/apcche.2004.0.991.0

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Different Roles of Metal Ions Both in the Formation of Amyloid-β Fibri and Superoxide

Nagami Hideto, Kishida Manabu, Umakoshi Hiroshi, Shimanouchi Toshinori, Kuboi Ryoichi

アジア・太平洋化学工学会議発表論文要旨集 2004 498 - 498 2004年

記述言語：英語出版者・発行元：公益社団法人化学工学会

The roles of metal ions both on the formation of fibril aggregation of amyloid-β-peptide (Aβ) and on the generation of superoxide by a metal-Aβ complex have been studied to clarify the relationship of both phenomena with the cause of Alzheimer's Disease (AD). Among the several metal ions, the addition of Zn(II) or of Cu(II) was found to increase the formation of Aβ fibril. Although Zn(II) promoted the Aβ fibril formation both at neutral and acidic pH levels, Cu(II) could induce the aggregation of Aβ only at acidic pH. In addition, hydrogen peroxide was generated at neutral pH in the absence of Aβ and Cu(II) beyond a stoichiometric amount of Cu(II) ion, suggesting the formation of a catalytic site that can react between Cu(II) and Cu(I). These results indicate that Zn(II) and Cu(II) at different pHs may each contribute to AD in different modes; the promotion of fibril aggregation (Zn(II) at every pH and Cu(II) at acidic pH) and the formation of superoxide (Cu(II) at neutral pH). The most characteristic aspect is the effect of pH on the promotion of aggregation and the formation of superoxide for the Cu(II)-Aβ system. The coordination structure of the Cu(II)-Aβ complex at neutral and acidic pHs was analyzed by using NMR spectroscopy, circular dichroism (CD), and ESR. The formation of a complex of histidine (His) residues and Cu(II) was found, and the coordination number of the His was suggested to be two at neutral pH and three at acidic pH. These results imply that the intramolecular coordination structure of the Aβ -Cu(II) complex at neutral pH could be converted to the intermolecular one by acidification. The pH variation thus led to a change in the complex structure of Cu(II)-Aβ. This finding suggests the important roles of pH to regulate two phenomena, i.e. the aggregation of Aβ (inhibition of superoxide formation) and the formation of superoxide from Cu(II)-Aβ complex.

DOI： 10.11491/apcche.2004.0.498.0

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Characterization of Stress-Response Behaviors of SOD/CAT on Liposome under Heat/Oxidative Stresses

Yoshimoto Noriko, Nagami Hideto, Umakoshi Hiroshi, Shimanouchi Toshinori, Kuboi Ryoichi

アジア・太平洋化学工学会議発表論文要旨集 2004 481 - 481 2004年

記述言語：英語出版者・発行元：公益社団法人化学工学会

A biological membrane is the frontier of the cells against the environmental reactive oxygen species (ROS) and is expected to play an important role in the antioxidant system corporately with antioxidant enzymes. The response behaviors of the antioxidant enzymes (superoxide dimutase (SOD) and catalase (CAT)) were investigated in the presence of artificial membrane, liposome, under the oxidative stress condition in order to obtain the fundamental information for the ROS scavenger system. The intracellular antioxidant enzymes, SOD and CAT, are known to make protective system against ROS. Although the SOD is known to catalyze the dismutation of ·O₂- to H₂O₂. The activity of SOD was lost thoroughly in the presence of H₂O₂ at high concentration (>5mM). The SDS-PAGE analysis showed that the SOD was fragmented into several small peptides. SOD has two -helix loops, which introduce the substrate ·O₂- to the activity center of SOD (Cu(II)). From the circular dichroism analysis of SOD in the presence of H₂O₂, the contents of α-helix in the SOD were found to decrease in corresponding with the inactivation of the SOD, suggesting that the conformation of the α-helix loops might be varied. However, the SOD was not inactivated in the presence of POPC liposomes while the contents of α-helix were decreased similarly in the case of that without liposome. Based on these results, the possibility to develop the ROS scavenger system using the SOD and CAT together with liposome was finally discussed.

DOI： 10.11491/apcche.2004.0.481.0

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3I10-2 ストレス条件下におけるアミロイド形成挙動に及ぼす脂肪酸修飾リポソームの共存効果(生物化学工学・分離精製工学,一般講演)

李奉局, 西村大佑, 馬越大, 島内寿徳, 久保井亮一

日本生物工学会大会講演要旨集 16 229 - 229 2004年

3I10-1 リポソームによる新規なバイオセンサーの設計・開発 : 水相中ストレス成分のモニタリングへの応用(生物化学工学・分離精製工学,一般講演)

佐々木勝司, 李奉局, 島内寿徳, 森田誠一, 久保井亮一

日本生物工学会大会講演要旨集 16 229 - 229 2004年

固定化リポリームクロストグラフィーによる生理活性物質の分離・分析招待査読

ぶんせき 11 636 - 642 2003年

Characterization of the surface properties of chitosanase under heat stress condition using aqueous two-phase systems and an immobilized-liposome sensor system

HS Jung, H Umakoshi, SY Son, T Shimanouchi, R Kuboi

SOLVENT EXTRACTION RESEARCH AND DEVELOPMENT-JAPAN 10 ( 10 ) 123 - 132 2003年

記述言語：英語出版者・発行元：JAPAN ASSOC SOLVENT EXTRACTION

The surface properties of chitosanase from Storeptomyces griseus were characterized under heat stress conditions by using aqueous two-phase systems and by using an immobilized-liposome sensor. The surface net (HFS) and local hydrophobicities (LH), were first characterized by using the poly (ethylene glycol) (PEG)/dextran 100-200k (Dex) aqueous two-phase system at various temperatures (25similar to50degreesC). The LH values of dithiothreitol (DTT)-treated chitosanase showed a maximum at a specific temperature (45degreesC) while the HFS values gradually increased with increasing temperature from 40 to 50degreesC. At the same temperature conditions as above, the surface properties of chitosanase were also analyzed by using a sensor tip immobilized-liposome electrode. Similarly in the case of the variation of LH but not HFS, the detected signal (maxima in electric current) was maximized at 45degreesC, and a linear correlation between the LH values and the signal values of the new immobilized-liposome sensor system was obtained. It was thus found that the new sensor system was effective for the characterization of the surface properties and LH values, of proteins, as well as the aqueous two-phase system method.

Web of Science

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1K13-5 人工 SOD リポソームの開発と酸化ストレスによる SOD 活性の制御

永見英人, 北浦武明, 島内寿徳, 馬越大, 久保井亮一

日本生物工学会大会講演要旨集 15 215 - 215 2003年

医薬品製造における晶析技術タンパク質の構造異常化によるストレス誘導型アミロイドーシス

島内寿徳, 馬越大, 久保井亮一

分離技術 32 ( 6 ) 347 - 351 2002年11月

Eraluation of Interaction between Liposome Membranes Induced by Stimuli Reponsire Polymers and Proteins

Journal of Bioscience and Bioengineering 93(5), 498-501/, 2002年

ストレス応答型バイオプロセスのためのストレスセンサの開発招待

鄭浩燮, 島内寿徳, 馬越大, 久保井亮一, 金廣, 櫻井芳昭, 夏川一輝

ケミカルエンジニヤリング 47 752 - 755 2002年

タンパク質の構造異常・アミロイドーシスとそのセンシング招待

島内寿徳, 佐々木勝司, 久保井亮一

ケミカルエンジニヤリング 47 748 - 751 2002年

Selective separation process of proteins based on the heat stress-induced translocation across phospholipid membranes

UMAKOSHI Hiroshi, SHIMANOUCHI Toshinori, KUBOI Ryoichi

Journal of chromatography. B, Biomedical sciences and applications 711 ( 1 ) 111 - 116 1998年6月

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