Interaction of Escherichia coli F1-ATPase with dicyclohexylcarbodiimide-binding polypeptide.
Antibody raised against the N,N'-dicyclohexylcarbodiimide (DCCD)-binding polypeptide of Escherichia coli bound to the cytoplasmic surface of the cell membrane. A weak reaction was seen with everted vesicles of the thermophile PS3. Rat-liver mitochondrial membranes did not react with the antibody. Reaction of the isolated DCCD-binding polypeptide with the antibody was prevented by oxidation of methionine residues or cleavage of the polypeptide with cyanogen bromide. Modification of the arginine residues of the DCCD-binding polypeptide did not affect interaction with the antibody. Purified F1-ATPase of E. coli bound to the isolated DCCD-binding polypeptide as shown by solid-phase radioimmune assay. Binding involved the alpha and/or beta subunits of F1 and the arginine residues of the polar central region of the DCCD-binding polypeptide. Our results are consistent with a looped arrangement of the DCCD-binding polypeptide in the membrane in which the carboxyl- and amino-terminal regions of the molecule are at the periplasmic surface and the polar central region, interacting with F1, is at the cytoplasmic surface of the cell membrane.
D000595:Amino Acid Sequence; D000936:Antigen-Antibody Complex; D002234:Carbodiimides / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D004024:Dicyclohexylcarbodiimide / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D007106:Immune Sera; D007700:Kinetics; D006180:Proton-Translocating ATPases / Q000378:metabolism*
D000936:Antigen-Antibody Complex; D002234:Carbodiimides; D002352:Carrier Proteins; D007106:Immune Sera; D004024:Dicyclohexylcarbodiimide; D006180:Proton-Translocating ATPases