A Catalytic Amino Acid and Primary Structure of Active Site inAspevgillus nigerα-Glucosidase
| Autor: | Masuhiro Takata, Seiya Chiba, Hirokazu Matsui, Haruhide Mori, Atsuo Kimura, Yukiharu Fukushi |
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| Rok vydání: | 1997 |
| Předmět: |
Tris
Stereochemistry Molecular Sequence Data Peptide Applied Microbiology and Biotechnology Biochemistry Analytical Chemistry Structure-Activity Relationship Residue (chemistry) chemistry.chemical_compound Amino Acid Sequence Enzyme Inhibitors Molecular Biology Peptide sequence chemistry.chemical_classification Aspartic Acid Binding Sites Sequence Homology Amino Acid biology Organic Chemistry Aspergillus niger Protein primary structure Active site alpha-Glucosidases General Medicine biology.organism_classification Amino acid Kinetics chemistry biology.protein Sequence Analysis Inositol Biotechnology |
| Zdroj: | Bioscience, Biotechnology, and Biochemistry. 61:1091-1098 |
| ISSN: | 1347-6947 0916-8451 |
| DOI: | 10.1271/bbb.61.1091 |
| Popis: | The catalytic amino acid residue of Aspergillus niger alpha-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 analyzing the sequence of peptide fragments prepared by several proteases. |
| Databáze: | OpenAIRE |
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