Identification of functional domains and critical residues in the adenosinetriphosphatase inhibitor protein of mitochondrial F0F1 ATP synthase.
Peptide segments of the inhibitor protein (IF1) of the F0F1 ATP synthase complex from bovine-heart mitochondria have been constructed by chemical synthesis. The IF1-(42-58)-peptide was equally effective as IF1 in inhibiting the ATPase activity of both the F0F1 complex in the mitochondrial membrane deprived of IF1 (SMP) and soluble F1. The IF1-(22-46)-peptide inhibited the ATPase activity in the soluble F1 but had no effect on either the ATPase activity or H+ conduction in SMP. Substitution of the His or Lys residues with Ala in the IF1-(42-58)-peptide decreased the inhibition of ATP hydrolysis. The inhibition exerted by the IF1-(42-58)-peptide on ATP hydrolysis in SMP exhibited a pH dependence, similar to that observed with IF1, which was lost upon replacement of His or Lys with Ala. In soluble F1, inhibition of ATP hydrolysis by IF1, the IF1-(42-58)-peptide and the IF1-(22-46)-peptide was pH dependent when F1 was first incubated with ATP. The IF1-(42-58)-peptide also caused inhibition of passive H+ conduction in SMP. This activity of the synthetic peptide was weaker, as compared to that of IF1, and practically unaffected by substitution of His or Lys with Ala. An antibody against the IF1-(42-58)-synthetic peptide stimulated ATP hydrolysis in the membrane-bound F0F1 complex with associated IF1 but was without effect on H+ conduction. An antibody against IF1 stimulated both processes.
Papa
S
S
Institute of Medical Biochemistry and Chemistry, University of Bari, Italy.
Zanotti
F
F
Cocco
T
T
Perrucci
C
C
Candita
C
C
Minuto
M
M
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002417:Cattle; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008929:Mitochondria, Heart / Q000201:enzymology; D008969:Molecular Sequence Data; D009154:Mutation; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011522:Protons; D000097795:ATPase Inhibitory Protein
D000255:Adenosine Triphosphate; D000906:Antibodies; D004791:Enzyme Inhibitors; D010446:Peptide Fragments; D011506:Proteins; D006180:Proton-Translocating ATPases; D011522:Protons; D000097795:ATPase Inhibitory Protein