Purification and characterization of the F/sub 1/-ATPase form Clostridium thermoaceticum
Journal Article
·
· J. Bacteriol.; (United States)
OSTI ID:5811395
The F/sub 1/ portion of the H/sup +/ -ATPase from Clostridium thermoaceticum was purified to homogeneity by solubilization at low ionic strength, ion-exchange chromatography, and gel filtration. The last indicated the M/sub r/ to be 370,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the pure enzyme revealed four bands with M/sub r/ corresponding to 60,000, 55,000, 37,000, and 17,000 in an apparent molar ratio of 3:3:1:1. The purified enzyme would bind to stripped membranes to reconstitute dicyclohexylcarbodiimide-sensitive ATPase activity. Phosphohydrolase activity, measured at 58/sup 0/C, was optimal at pH 8.5. In the presence of a 1 mM excess of Mg/sup 2 +/ over the concentration of ATP, the K/sub m/ for ATP was 0.4 mM, and the V/sub max/ was 6.7 ..mu..mol min /sup -1/ mg/sup -1/. Unlike the membrane-bound F/sub 1/F/sub 0/ complex, the F/sub 1/-ATPase was relatively insensitive to the inhibitors dicyclohexylcarbodiimide and tributyltin chloride. Both the complex and the F/sub 1/-ATPase were inhibited by quercetin, azide, 7-chloro-4-nitro-benz-2-oxa-1,3-diazole, and free magnesium, and both were stimulated by primary alcohols and sulfite. In whole cells, the F/sub 1/F/sub 0/-ATPase catalyzed the synthesis of ATP in response to a pH gradient.
- Research Organization:
- Univ. of Georgia, Athens
- DOE Contract Number:
- AS09-79ER10499
- OSTI ID:
- 5811395
- Journal Information:
- J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 165:1; ISSN JOBAA
- Country of Publication:
- United States
- Language:
- English
Similar Records
Characterization of the ATP synthase of Propionigenium modestum as a primary sodium pump
Generation of energy during CO/sub 2/ fixation in acetogenic bacteria
Identification of a vanadate-sensitive, membrane-bound ATPase in the archaebacterium methanococcus voltae
Journal Article
·
Tue Sep 20 00:00:00 EDT 1988
· Biochemistry; (USA)
·
OSTI ID:6897679
Generation of energy during CO/sub 2/ fixation in acetogenic bacteria
Thesis/Dissertation
·
Wed Dec 31 23:00:00 EST 1986
·
OSTI ID:6281078
Identification of a vanadate-sensitive, membrane-bound ATPase in the archaebacterium methanococcus voltae
Journal Article
·
Tue Sep 01 00:00:00 EDT 1987
· J. Bacteriol.; (United States)
·
OSTI ID:6062083