Induced versus pre-existing asymmetry models for the half-of-the-sites reactivity effect in bovine liver uridine diphosphoglucose dehydrogenase
| Autor: | David Sidney Feingold, Cardamone Jj, James S. Franzen, Paul S. Marchetti, J Ashcom |
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| Rok vydání: | 1980 |
| Předmět: |
chemistry.chemical_classification
Binding Sites biology Chemistry Stereochemistry Protein Conformation Dehydrogenase Iodoacetates General Medicine Uridine Diphosphate Glucose Dehydrogenase Enzyme assay Catalysis Iodoacetamide Microscopy Electron Enzyme Liver Naphthalenesulfonates Covalent bond Thiol biology.protein Animals Reactivity (chemistry) Carbohydrate Dehydrogenases Cattle NAD+ kinase |
| Zdroj: | Biochimica et biophysica acta. 614(2) |
| ISSN: | 0006-3002 |
| Popis: | Half-of-the-sites reactivity of the catalytic site thiol groups of UDPglucose dehydrogenase (UDPglucose:NAD+ 6-oxidoreductase, EC 1.1.1.22) can be ascribed either to the induction of conformational asymmetry following derivatization of one half of the subunits or to intrinsic conformational differences in the subunits of the native enzyme. If the half-sites reactivity behavior is due to induction effects, the magnitude of the induction could be expected to depend on the nature of the covalent modification. On the other hand, if the half-sites reactivity behavior is due to pre-existing asymmetry and there is no communication between catalytic centers, the properties of unmodified sub-units should be independent of the nature of the covalent derivative introduced on the modified subunits. According to the induced asymmetry hypothesis, the catalytic activity of half-sites modified enzyme might be different for different covalent modifications, whereas for the rigid pre-existing asymmetry hypothesis the catalytic activity of half-sites modified enzyme should be the same regardless of the modifying group. During the course of catalytic site thiol group modification by a number of thiol specific reagents, the loss of enzyme activity was equivalent to the degree of modification for most of the reagents employed. However, with iodoacetate and 5-(iodoacetamidoethyl)aminonaphthalene-1-sulfonic acid, half-sites modification of UDPglucose dehydrogenase reduced catalytic activity by 58 and 78%, respectively, of the initial activity. These observations are consistent with a model in which there is communication between catalytic sites. Electron microscopy shows that the six subunits of UDPglucose dehydrogenase are arranged as a hexagonal planar ensemble. |
| Databáze: | OpenAIRE |
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