Nitrate assimilation in Neurospora crassa: Enzymatic and immunoblot analysis of wild-type and nit mutant protein products in nitrate-induced and glutamine-repressed cultures
| Autor: | Reginald H. Garrett, Barry K. Hurlburt |
|---|---|
| Rok vydání: | 1988 |
| Předmět: |
Glutamine
Nitrogen assimilation Nitrate reductase Nitrate Reductase Neurospora Neurospora crassa chemistry.chemical_compound Nitrate Nitrate Reductases Genetics Molecular Biology Immunosorbent Techniques chemistry.chemical_classification Nitrates biology Wild type biology.organism_classification Nitrite reductase Molecular biology Enzyme chemistry Biochemistry Mutation Enzyme Repression Apoproteins |
| Zdroj: | Molecular and General Genetics MGG. 211:35-40 |
| ISSN: | 1432-1874 0026-8925 |
| DOI: | 10.1007/bf00338390 |
| Popis: | The nitrate assimilatory pathway in Neurospora crassa is composed of two enzymes, nitrate reductase and nitrite reductase. Both are alpha 2 type homodimers. Enzyme-bound prosthetic groups mediate the electron transfer reactions which reduce inorganic nitrate to an organically utilizable form, ammonium. One, a molybdenum-containing cofactor, is required by nitrate reductase for both enzyme activity and holoenzyme assembly. Three modes of regulation are imposed on the expression of nitrate assimilation, namely: nitrogen metabolite repression, nitrate induction and autogenous regulation by nitrate reductase. In this study, nitrocellulose blots of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) resolved proteins from crude extracts of the wild type and specific nitrate-nonutilizing (nit) mutants were examined for material cross-reactive with antibodies against nitrate reductase and nitrite reductase. The polyclonal antibody preparations used were rendered monospecific by reverse affinity chromatography. Growth conditions which alter the regulatory response of the organism were selected such that new insight could be made into the complex nature of the regulation imposed on this pathway. The results indicate that although nitrate reductase and nitrite reductase are coordinately expressed under specific nutritional conditions, the enzymes are differentially responsive to the regulatory signals. |
| Databáze: | OpenAIRE |
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