The inhibitor protein (IF1) promotes dimerization of the mitochondrial F1F0-ATP synthase.
The effect of increased expression or reconstitution of the mitochondrial inhibitor protein (IF1) on the dimer/monomer ratio (D/M) of the rat liver and bovine heart F1F0-ATP synthase was studied. The 2-fold increased expression of IF1 in AS-30D hepatoma mitochondria correlated with a 1.4-fold increase in the D/M ratio of the ATP synthase extracted with digitonin as determined by blue native electrophoresis and averaged densitometry analyses. Removal of IF1 from rat liver or bovine heart submitochondrial particles increased the F1F0-ATPase activity and decreased the D/M ratio of the ATP synthase. Reconstitution of recombinant IF1 into submitochondrial particles devoid of IF1 inhibited the F1F0-ATPase activity by 90% and restored partially the D/M ratio of the whole F1F0 complex as revealed by blue native electrophoresis and subsequent SDS-PAGE or glycerol density gradient centrifugation. Thus, the inhibitor protein promotes or stabilizes the dimeric form of the intact F1F0-ATP synthase. A possible location of the IF1 protein in the dimeric structure of the rat liver F1F0 complex is proposed. According to crystallographic and electron microscopy analyses, dimeric IF1 could bridge the F1-F1 part of the dimeric F1F0-ATP synthase in the inner mitochondrial membrane.
García
José J
JJ
Departamento de Bioquímica and Cardiovascular Disease's Genomic and Proteomic Study Group, Instituto Nacional de Cardiología Ignacio Chavez, Juan Badiano 1, Col. Sección XVI, México, DF, Mexico 14080. jjgarcia_trejo@yahoo.com
Morales-Ríos
Edgar
E
Cortés-Hernandez
Paulina
P
Rodríguez-Zavala
José S
JS
D000818:Animals; D002417:Cattle; D019281:Dimerization; D008114:Liver Neoplasms, Experimental; D008930:Mitochondria, Liver / Q000378:metabolism*; D008958:Models, Molecular; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D051381:Rats; D013367:Submitochondrial Particles / Q000378:metabolism*; D000097795:ATPase Inhibitory Protein
D011506:Proteins; D006180:Proton-Translocating ATPases; D000097795:ATPase Inhibitory Protein