Extraction of mitochondrial protein-lipid complexes into organic solvents: an approach to study the interaction between the ATPase and the mitochondrial ATPase-inhibitor protein.
Protein-lipid complexes were transferred directly from mitochondria and submitochondrial particles into hexane and ether. The protein-lipid residue left after solvent removal from these extracts was used to form liposomes which display low-temperature-resistant ATPase activity. Centrifugation experiments indicate that the ATPase activity is associated to the vesicles. Most of the F1-ATPases appear to be accessible to the external water phase of the liposomes. The ATPase activity of these particles was insensitive to dicyclohexylcarbodiimide and oligomycin. Incubation of these vesicles at room temperature activated (4--10-fold) the ATPase through a process that is partially sensitive to phenylmethylsulfonyl fluoride. The results with purified ATPase-inhibitor protein and (F1--ATPase)-inhibitor complex indicate that the activation process in the liposomes is due to the abolition of the inhibitory action of the inhibitor protein bound to a large fraction of the extracted ATPases. Liposomes prepared from hexane extracts obtained from submitochondrial particles having different levels of ATPase activity displayed an activation ratio which correlated with the number of ATPases that are inhibited by the inhibitor protein in the submitochondrial particles. The extraction of mitochondrial ATPase and its incorporation into liposomes followed by activity measurements may be used to judge the number of ATPases that in a given preparation contain the inhibitor protein in its inhibiting site.
D000251:Adenosine Triphosphatases / Q000302:isolation & purification*; D000818:Animals; D002417:Cattle; D008055:Lipids / Q000302:isolation & purification*; D008081:Liposomes / Q000378:metabolism; D008929:Mitochondria, Heart / Q000378:metabolism*; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D012997:Solvents; D013347:Subcellular Fractions / Q000378:metabolism; D000097795:ATPase Inhibitory Protein
D000251:Adenosine Triphosphatases; D008055:Lipids; D008081:Liposomes; D011506:Proteins; D012997:Solvents; D000097795:ATPase Inhibitory Protein