Lipid protein interactions in mitochondria. VII. A comparison of the effects of lipid removal and lipid perturbation of the kinetic properties of mitochondrial ATPase.
We investigated the kinetics of mitochondrial ATPase in bovine heart mitochondria and submitochondrial particles upon treatment with phospholipase A2, or upon addition of n-butanol to perturb the lipid protein interactions. The changes observed are the following: (1) Lipid removal or perturbation with butanol is accompanied by loss of ATPase activity with decrease of both V and of the KM for ATP. (2) There are changes of activation energy of ATPase activity at temperatures above the discontinuity normally observed for membrane-bound enzymes in mitochondria. In particular, butanol abolishes the discontinuity, and induces a constant activation energy of about 32 kcal/mol in the range 8--37 degrees C. (3) Butanol modifies the pH dependence of ATPase shifting the pH optimum from around 10 to less alkaline values. The optimum for Mg2+ concentrations is increased by the solvent. (4) Treatment with phospholipase A2 results in a removal of oligomycin-sensitive ATPase, whereas butanol addition prevents oligomycin inhibition of ATPase. (5) In beef heart mitochondria, a spin-labelled analog of the inhibitor, dicyclohexyl carbodiimide, did not show any change in environment upon butanol addition, unlike that found in mitochondria from Saccharomyces cerevisiae.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D000440:Butanols / Q000494:pharmacology; D002417:Cattle; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008928:Mitochondria / Q000201:enzymology*; D008929:Mitochondria, Heart / Q000187:drug effects / Q000201:enzymology*; D009840:Oligomycins / Q000494:pharmacology; D010740:Phospholipases; D010743:Phospholipids / Q000502:physiology*; D013367:Submitochondrial Particles / Q000187:drug effects / Q000201:enzymology*; D013816:Thermodynamics
D000440:Butanols; D009840:Oligomycins; D010743:Phospholipids; D010740:Phospholipases; D000251:Adenosine Triphosphatases; D008274:Magnesium