Formation in Vivo, Purification and Crystallization of a Complex of the γ and ε Subunits of the F0F1-ATPase of Escherichia coli
| Autor: | Robert G. Solomon, Cromer Ba, Graeme B. Cox, Guss Jm, Harvey I, Dianne C. Webb, Jeffrey Pd |
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| Rok vydání: | 1993 |
| Předmět: |
Stereochemistry
Recombinant Fusion Proteins ATPase Protein subunit medicine.disease_cause Thrombin Structural Biology In vivo Escherichia coli medicine Molecular Biology Glutathione Transferase chemistry.chemical_classification biology Chromatography Ion Exchange biology.organism_classification Enterobacteriaceae Fusion protein Proton-Translocating ATPases Enzyme Biochemistry chemistry biology.protein Electrophoresis Polyacrylamide Gel Crystallization Plasmids medicine.drug |
| Zdroj: | Journal of Molecular Biology. 229:1159-1162 |
| ISSN: | 0022-2836 |
| DOI: | 10.1006/jmbi.1993.1113 |
| Popis: | A complex comprising the epsilon subunit of Escherichia coli F1-ATPase (ECF1-ATPase) and a glutathione-S-transferase gamma subunit (of ECF1-ATPase) fusion protein was formed in vivo and purified from cell extracts by binding to glutathione-agarose beads. The glutathione-S-transferase was released from the complex by digestion with thrombin and the gamma/epsilon complex purified by cation-exchange chromatography. Crystals of the complex were grown by vapour diffusion using PEG8000 as precipitant. The crystals are orthorhombic, space-group P2(1)2(1)2 with a = 161.9 A, b = 44.1 A and c = 63.4 A. The volume of the asymmetric unit is consistent with the presence of a complex of one gamma subunit and one epsilon subunit. |
| Databáze: | OpenAIRE |
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