Inactivation of the mitochondrial ATPase inhibitor protein by chemical modification with diethylpyrocarbonate.
Modification of histidine residue(s) by diethylpyrocarbonate treatment of submitochondrial particles obtained by sonication results in inhibition of ATPase activity and stimulation of oligomycin-sensitive H+ conduction. The inhibition of the ATPase (EC 3.6.1.3) activity persisted in F1 isolated from diethylpyrocarbonate-treated submitochondrial particles, which exhibited the absorbance spectrum of modified histidine. Thus the inhibition of the ATPase activity results from histidine modification in F1 subunits. Removal of the natural inhibitor protein from submitochondrial particles resulted in stimulation of proton conduction. After removal of F1 inhibitor protein from the particles the stimulatory effect exerted by diethylpyrocarbonate treatment on proton conduction was lost. Reconstitution experiments showed that purified F1 inhibitor protein lost, after histidine modification, its capacity to inhibit the ATPase activity and proton conduction. These observations show that the stimulation of proton conduction by the ATPase complex effected by diethylpyrocarbonate treatment results from histidine modification in F1 inhibitor protein.
D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D006639:Histidine; D008929:Mitochondria, Heart / Q000201:enzymology*; D011506:Proteins / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors; D011522:Protons; D013053:Spectrophotometry; D013367:Submitochondrial Particles / Q000187:drug effects / Q000201:enzymology; D000097795:ATPase Inhibitory Protein
D004047:Diethyl Pyrocarbonate; D004492:Edetic Acid; D006639:Histidine; D011506:Proteins; D006180:Proton-Translocating ATPases; D011522:Protons; D000097795:ATPase Inhibitory Protein