Purification and characteristics of hydrophobic membrane protein(s) required for DCCD sensitivity of ATPase in mycobacterium phlei
✍ Scribed by Cohen, Natalie S. ;Lee, Soon-Ho ;Brodie, Arnold F.
- Publisher
- Wiley (John Wiley & Sons)
- Year
- 1978
- Tongue
- English
- Weight
- 458 KB
- Volume
- 8
- Category
- Article
- ISSN
- 0091-7419
No coin nor oath required. For personal study only.
✦ Synopsis
The energy-transducing N,N'-dicyclohexylcarbodiimide-sensitive (DCCDsensitive) ATPase complex consists of two parts, a soluble catalytic protein (F, ), and an intrinsic membrane protein (Fo). The bacterial coupling factor complex, BCF,-BCF,, has recently been purified from Mycobacterium phlei, and used to reconstitute oxidative phosphorylation in detergent-extracted membranes. The BCFo moiety has been purified by being recovered from the purified BCFo-BCF, complex by affinity chromatography. BCF, is a lipoprotein or lipoprotein complex with an approximate molecular weight of 60,000. The preparation contained 0.15 mg of phospholipid per milligram protein. There appear t o be three polypeptides, with approximate molecular weights of 24,000, 18,000, and 8,000 as determined by sodium dodecylsulfate acrylamide gel electrophoresis. Purified BCF, conferred DCCD sensitivity on a purified BCF, preparation. Reconstitution of oxidative phosphorylation was achieved after incubation of detergent-extracted membranes with purified BCF, and purified BCFl.