Purification of 5-Aminolaevulinate Synthase from Liver Mitochondria of Chick Embryo
| Autor: | Brian K. May, J.D. Brooker, Gopesh Srivastava, W.H. Elliott, Iain A. Borthwick |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
chemistry.chemical_classification
Chromatography ATP synthase Immunoprecipitation Chromatofocusing Sodium chemistry.chemical_element Mitochondria Liver Chick Embryo Biology Biochemistry Molecular biology Molecular Weight Enzyme Affinity chromatography chemistry biology.protein Animals Electrophoresis Polyacrylamide Gel Specific activity Polyacrylamide gel electrophoresis 5-Aminolevulinate Synthetase |
| Zdroj: | European Journal of Biochemistry. 129:615-620 |
| ISSN: | 1432-1033 0014-2956 |
| Popis: | 5-Aminolaevulinate synthase from chick-embryo liver mitochondria has, for the first time, been purified to homogeneity in its native non-degraded form by molecular sieve chromatography, chromatofocusing and affinity chromatography. The enzyme has a minimum molecular weight of 68000 as determined by sodium dodecylsulphate/polyacrylamide gel electrophoresis and a specific activity of 35000 units/mg of protein. This result conflicts with the previous report of Whiting, M. J. and Granick, G. [(1976) J. Biol. Chem 251, 1340–1346] that the chick embryo enzyme has a molecular weight of 49000. We show here that the purified form can be degraded proteolytically to a smaller form of molecular weight around 50000 while retaining full enzymatic activity. It seem evident, therefore, that the enzyme isolated by Whiting & Granick (1976) was degraded. We have further established by pulse-labelling studies and immunoprecipitation that the enzyme isolated by our new and rapid procedure has the same minimum molecular weight as that which exists in vivo. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|