Peptide analogs of the beef heart mitochondrial F1-ATPase inhibitor protein.
Peptide analogs which correspond to the conserved region of the natural ATPase inhibitor protein from beef heart, Candida utilis, and Saccharomyces cerevisiae mitochondria were synthesized by solid-phase methodologies and tested for ATPase inhibitory activity. These peptides were found to be potent inhibitors of F1-ATPase-catalyzed ATP hydrolysis in acidic reaction media, having I50 values of 1.1 +/- 0.4 microM, 10 +/- 5 microM, and 48 +/- 19 microM, respectively. These results closely match those obtained for the naturally occurring inhibitor proteins. Additional peptides that correspond to the beef heart beta-subunit near the binding site of the beef heart inhibitor protein and that possess a substantial homology with the conserved region of the inhibitor protein were synthesized. Several of these peptides were found to be inhibitors of the ATPase activity. The best inhibitor, with an I50 value of 20 +/- 3 microM, was the peptide resembling the beef heart beta-subunit comprising amino acids 394-413. This peptide most closely resembles the peptides derived from the conserved region of the inhibitor protein. The insertion of five glycine residues between the charge clusters in the beta-394-413 peptide resulted in a peptide which was able to stimulate the hydrolysis of ATP.
Stout
J S
JS
Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610-0245.
Partridge
B E
BE
Dibbern
D A
DA
Schuster
S M
SM
D000595:Amino Acid Sequence; D000818:Animals; D002175:Candida / Q000737:chemistry; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008928:Mitochondria / Q000737:chemistry; D008929:Mitochondria, Heart / Q000737:chemistry*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors*; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D000097795:ATPase Inhibitory Protein
D010446:Peptide Fragments; D011506:Proteins; D006180:Proton-Translocating ATPases; D000097795:ATPase Inhibitory Protein