Effects of naturally occurring polyols and urea on mitochondrial F0F1ATPase.
We show that urea inhibits the ATPase activity of MgATP submitochondrial particles (MgATP-SMP) with Ki = 0.7 M, probably as a result of direct interaction with the structure of F0F1-ATPase. Counteracting compounds (sorbitol, mannitol or inositol), despite slightly (10-20%) inhibiting the ATPase activity, also protect the F0F1-ATPase against denaturation by urea. However, this protection was only observed at low urea concentrations (less than 1.5 M), and in the presence of three polyols, the Ki for urea shift from 0.7 M to 1.2 M. Urea also increases the initial activation rate of latent MgATP-SMP in a dose-dependent-manner. However, when the particles (0.5 mg/ml) were preincubated in the presence of 1 M, 2 M or 3 M urea, a decrease in the activation level occurred after 1 h, 30 and 10 min, respectively. At high MgATP-SMP concentration (3 mg/ml) a decrease in activation was observed after 2 h, 1 h and 20 min, respectively. These data indicate that the effect of urea on the activation of MgATP-SMP depends on time, urea and protein concentrations. It was also observed that polyols suppress the activation of latent MgATP-SMP in a dose-dependent manner, and protect the particles against urea denaturation during activation. We suppose that a decrease in membrane mobility promoted by interactions of polyols with phospholipids around the F0F1-ATPase may also increase the compactation of protein structure, explaining the inhibition of natural inhibitor protein of ATPase (IF1) release and the activation of the enzyme.
Lemos
A P
AP
Departamento de Bioquímica Médica, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Ilha do Fundão, RJ, Brazil.
Peres-Sampaio
C E
CE
Guimarães-Motta
H
H
Silva
J L
JL
Meyer-Fernandes
J R
JR
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002417:Cattle; D007294:Inositol / Q000494:pharmacology; D007700:Kinetics; D008353:Mannitol / Q000494:pharmacology; D008929:Mitochondria, Heart / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D013012:Sorbitol / Q000494:pharmacology; D013367:Submitochondrial Particles / Q000201:enzymology*; D013402:Sugar Alcohols / Q000494:pharmacology*; D014508:Urea / Q000494:pharmacology*
D013402:Sugar Alcohols; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D008353:Mannitol; D007294:Inositol; D013012:Sorbitol; D000255:Adenosine Triphosphate; D014508:Urea; D006180:Proton-Translocating ATPases