Proteolytic cleavage of soybean Bowman-Birk inhibitor monitored by means of high-performance capillary electrophoresis. Implications for the mechanism of proteinase inhibitors
| Autor: | Peter Flecker, Jochen Uebe, Klaus K. Unger, Birte Jensen, Young-Mi Kim, Manfred Gey |
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| Rok vydání: | 1996 |
| Předmět: |
chemistry.chemical_classification
Binding Sites Chromatography biology Chemistry Hydrolysis Molecular Sequence Data Biophysics Electrophoresis Capillary Active site Cleavage (embryo) Biochemistry Catalysis Protein Structure Tertiary Kinetics Electrophoresis Reaction rate constant Enzyme Capillary electrophoresis Enzyme inhibitor biology.protein Amino Acid Sequence Trypsin Inhibitor Bowman-Birk Soybean |
| Zdroj: | Journal of Biochemical and Biophysical Methods. 33:171-185 |
| ISSN: | 0165-022X |
| Popis: | The hydrolysis of the soybean Bowman-Birk inhibitor in the presence of catalytic amounts of bovine trypsin and the formation of the non-covalent enzyme-inhibitor complex with an equimolar amount of enzyme are monitored by means of high-performance capillary electrophoresis (HPCE). The inhibitor is cleaved in the trypsin-reactive and more slowly in the chymotrypsin-reactive subdomain. HPCE proves itself as the only reliable analytical tool to monitor these reactions in clear contrast to classical electrophoretic, chromatographic and enzymatic methods. The most efficient separation of the intact and the two active site cleaved forms of the inhibitor was achieved in borate buffer at pH 10.0. The pH dependence of the rate constant and the final extent of hydrolysis reveal the stability of the enzyme inhibitor complex as a central aspect of the mechanism of proteinase inhibitors. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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