A Catalytic Amino Acid and Primary Structure of Active Site in Aspergillus niger α-Glucosidase
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概要
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The catalytic amino acid residue of Aspergillus niger α-glucosidase (ANGase) was identified by modification with conduritol B epoxide (CBE), a mechanism-based irreversible inactivator. The inactivation by CBE followed pseudo-first order kinetics. The interaction of CBE and ANGase conformed to a model with a reversible enzyme-inhibitor complex formed before covalent inactivation. A competitive inhibitor, Tris, decreased the inactivation rate. The incorporation of one mole of CBE per mole of ANGase was completely abolished the enzyme activity. A dissociated carboxyl group (-COO-) in the active site was suggested to attack the C-1 of CBE. ANGase was composed of two subunits (P1 and P2), of which P2 was modified by CBE. The labelled residue was included in a peptide (LY3) that was obtained from Lys-C protease digestion of CBE-bound P2. The sequence analysis of CBE-labelled LY3 showed that an Asp was the modified residue, that is, one of the catalytic amino acid residues of ANGase. The primary structure of LY3 was determined by analyiing the sequence of peptide fragments prepared by several proteases.
- 社団法人日本農芸化学会の論文
- 1997-07-23
著者
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森 春英
北海道大学大学院農学研究科応用生命科学専攻
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Takata Masuhiro
Department Of Applied Bioscience Faculty Of Agriculture Hokkaido University
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Matsui Hirokazu
Department of Bioscience and Chemistry, Faculty of Agriculture, Hokkaido University
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千葉 誠哉
Graduate School Of Agriculture Hokkaido University
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木村 淳夫
北海道大学大学院農学研究院応用生命科学部門
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Chiba Seiya
Department Of Agricultural Chemistry Faculty Of Agriculture Hokkaido University
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Chiba Seiya
Division Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Fukushi Yukiharu
Departments of Applied Bioscience Hokkaido University
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Matsui H
Division Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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MORI Haruhide
Department of Applied Bioscience, Graduate School of Agriculture, Hokkaido University
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KIMURA Atsuo
Department of Applied Bioscience, Graduate School of Agriculture, Hokkaido University
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Matsui Hirokazu
Dept. Biosci. Chem. Fac. Agric. Hokkaido Univ.
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Matsui Hirokazu
Department Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Fukushi Yukiharu
Department Of Applied Bioscience Faculty Of Agriculture Hokkaido University
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Kobayasi Kazuhiro
Department Of Agriculture Faculty Of Life And Environmental Science Shimane University
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Kimura Atsuo
Department Of Agricultural Chemistry Faculty Of Agriculture Hokkaido University
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Kimura Atsuo
Hokkaido University
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MATSUI Hirokazu
Department of Agricultural Chemistry, Faculty of Agriculture, Hokkaido University
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FUKUSHI Yukiharu
Department of Agricultural Chemistry, Faculty of Agriculture, Hokkaido University
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