Purification and properties of the latent F1-ATPase of Micrococcus lysodeikticus
| Autor: | Michael Huberman, Milton R.J. Salton |
|---|---|
| Rok vydání: | 1979 |
| Předmět: |
Adenosine Triphosphatases
Serine protease Gel electrophoresis Protease Chromatography biology Molecular mass Macromolecular Substances Chemistry medicine.medical_treatment ATPase Biophysics Cell Biology Biochemistry Hydrolysate Micrococcus Molecular Weight Oxidative Phosphorylation Coupling Factors medicine biology.protein Ultracentrifuge Amino Acids Immunoelectrophoresis Polyacrylamide gel electrophoresis |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Bioenergetics. 547:230-240 |
| ISSN: | 0005-2728 |
| DOI: | 10.1016/0005-2728(79)90006-9 |
| Popis: | The latent coupling factor (F1)-ATPase of Micrococcus lysodeikticus has been purified to homogeneity as determined by a number of criteria including, nondenaturing polyacrylamide gel electrophoresis, crossed immunoelectrophoresis and analytical ultracentrifugation. By inclusion of 1 mM phenylmethyl sulfonyl fluoride, a serine protease inhibitor, in the shock-wash step of release of F1 from the membranes, the spontaneous activation of both crude and purified ATPase by endogenous membrane protease(s) can be prevented, thereby yielding a highly latent ATPase preparation. Equilibrium ultracentrifugation of the latent ATPase gave a molecular weight of 400 000. The ATPase contained five different subunits alpha, beta, gamma, delta, and espsilon and their molecular weights determined by SDS-polyacrylamide gel electrophoresis were 60 000, 54 000, 37 000, 27 000 and 9000, respectively. The subunit composition was determined with 14C-labelled, F1-ATPase prepared from cells grown on medium containing [U-14C]-labelled algal protein hydrolysate. Within the limitations of this method the results tentatively suggest a subunit composition of 3 : 3 : 1 : 1 : 3. |
| Databáze: | OpenAIRE |
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