Clostridium pasteurianum F1Fo ATP synthase: operon, composition, and some properties.
The atp operon encoding F1Fo ATP synthase in the fermentative obligate anaerobic bacterium Clostridium pasteurianum was sequenced. It consisted of nine genes arranged in the order atpI(i), atpB(a), atpE(c), atpF(b), atpH(delta), atpA(alpha), atpG(gamma), atpD(beta), and atpC(epsilon), which was identical to that found in many bacteria. Reverse transcription-PCR confirmed the presence of the transcripts of all nine genes. The amount of ATPase activity in the membranes of C. pasteurianum was low compared to what has been found in many other bacteria. The F1Fo complexes solubilized from membranes of C. pasteurianum and Escherichia coli had similar masses, suggesting similar compositions for the F1Fo complexes from the two bacteria. Western blotting experiments with antibodies raised against the purified subunits of F1Fo detected the presence of eight subunits, alpha, beta, gamma, delta, epsilon, a, b, and c, in the F1Fo complex from C. pasteurianum. The F1Fo complex from C. pasteurianum was activated by thiocyanate, cyanate, or sulfhydryl compounds; inhibited by sulfite, bisulfite, or bicarbonate; and had tolerance to inhibition by dicyclohexylcarbodiimide. The target of thiol activation of the F1Fo complex from C. pasteurianum was F1. Thiocyanate and sulfite were noncompetitive with respect to substrate Mg ATP but competitive with respect to each other. The F1 and Fo parts of the F1Fo complexes from C. pasteurianum and E. coli bound to each other, but the hybrid F1Fo complexes were not functionally active.
Das
Amaresh
A
Center for Biological Resource Recovery, University of Georgia, Athens, Georgia 30602, USA. adas@arches.uga.edu
Ljungdahl
Lars G
LG
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001639:Bicarbonates / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D003013:Clostridium / Q000201:enzymology* / Q000235:genetics; D003485:Cyanates / Q000494:pharmacology; D003902:Detergents / Q000737:chemistry; D004492:Edetic Acid / Q000737:chemistry; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D025261:Mitochondrial Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009876:Operon; D021122:Protein Subunits; D012333:RNA, Messenger / Q000378:metabolism; D012995:Solubility; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013447:Sulfites / Q000494:pharmacology; D013861:Thiocyanates / Q000494:pharmacology; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D001639:Bicarbonates; D003485:Cyanates; D003902:Detergents; D004791:Enzyme Inhibitors; D021122:Protein Subunits; D012333:RNA, Messenger; D013438:Sulfhydryl Compounds; D013447:Sulfites; D013861:Thiocyanates; D004492:Edetic Acid; C474451:F1F0-ATP synthase; D025261:Mitochondrial Proton-Translocating ATPases; C031760:thiocyanate; C042345:hydrogen sulfite