Chemical modification of the F0 part of the ATP synthase (F1F0) from Escherichia coli. Effects on proton conduction and F1 binding.
The purified F0 part of the ATP synthase complex from Escherichia coli was incorporated into liposomes and chemically modified by various reagents. The modified F0-liposomes were assayed for H+ uptake and, after reconstitution with F1, for total and dicyclohexylcarbodiimide-sensitive ATPase activity. The water-soluble carbodiimide, 1-ethyl-3-(-3-dimethylaminopropyl)carbodiimide methiodide, (1.2 mM), inhibited H+ uptake to a great extent. Binding of F1 was almost unaffected, but the hydrolysis of ATP was uncoupled from H+ transport. This is reflected by the inhibition of dicyclohexylcarbodiimide-sensitive ATPase activity. Woodward's reagent K, N-ethyl-5-phenylisoxazolium-3'-sulfonate, inhibited both H+ uptake and total ATPase activity. Modification of arginine residues by phenylglyoxal (20 mM) was followed by inhibition of the F1 binding activity by 80% of the control. H+ translocation was reduced to 70%. Diethylpyrocarbonate (3 mM) exhibited a strong inhibiting effect on H+ uptake but not on F1 binding. Modification of tyrosine (by tetranitromethane) as well as lysine residues (by succinic anhydride) did not affect F0 functions. From the data presented we conclude that carboxyl-groups, different from the dicyclohexylcarbodiimide-binding site, are involved in H+ translocation through F0 and, in part, in the functional binding of F1. Furthermore, for the latter function, also arginine residues seem to be important. The role of histidine residues remains unclear at present.
D025181:ATP Synthetase Complexes; D001120:Arginine; D055598:Chemical Phenomena; D002621:Chemistry; D004047:Diethyl Pyrocarbonate; D004926:Escherichia coli / Q000201:enzymology*; D008081:Liposomes; D008239:Lysine; D009097:Multienzyme Complexes / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism*; D011485:Protein Binding; D011522:Protons; D014443:Tyrosine
D008081:Liposomes; D009097:Multienzyme Complexes; D010446:Peptide Fragments; D011522:Protons; D014443:Tyrosine; D001120:Arginine; D010770:Phosphotransferases; D025181:ATP Synthetase Complexes; D008239:Lysine; D004047:Diethyl Pyrocarbonate