Temperature effects on malonyl-CoA inhibition of carnitine palmitoyltransferase I.
Malonyl-CoA inhibition of hepatic mitochondrial carnitine palmitoyltransferase I and malonyl-CoA binding were measured at temperatures ranging from 0 degrees C to 37 degrees C. Protease treatment of mitochondria resulted in greatly diminished malonyl-CoA binding, indicating that the method used detected malonyl-CoA binding sites located on the outer surface of the mitochondrial outer membrane as expected. The apparent Ki for malonyl-CoA inhibition was found to increase with increasing temperature. Arrhenius plots for the initial velocity of the enzymatic reaction and for the Ki for malonyl-CoA both indicated a transition temperature between 20 and 25 degrees C with the transition for the malonyl-CoA interaction being more pronounced. Total specific binding of malonyl-CoA to mitochondrial proteins increased with increasing temperature, and Kd values decreased. The opposite effect of temperature on Kd values and Ki values was surprising because it was expected that these equilibrium constants would be identical. These observations indicate that Kd values for malonyl-CoA binding and Ki values for inhibition of carnitine palmitoyltransferase I by malonyl-CoA represent two significantly different binding phenomena. These data suggest that either: (a) malonyl-CoA binding measurements are unrelated to malonyl-CoA inhibition, or (b) inhibition of carnitine palmitoyltransferase I by malonyl-CoA involves more complex relationships than binding of malonyl-CoA to a single protein.
Kashfi
K
K
Department of Pharmacology, College of Medicine, University of Tennessee-Memphis 38163, USA.
Cook
G A
GA
D000818:Animals; D002334:Carnitine O-Palmitoyltransferase / Q000037:antagonists & inhibitors*; D003080:Cold Temperature; D010450:Endopeptidases; D007700:Kinetics; D008297:Male; D008316:Malonyl Coenzyme A / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology*; D008930:Mitochondria, Liver / Q000201:enzymology* / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D013696:Temperature
D008316:Malonyl Coenzyme A; D002334:Carnitine O-Palmitoyltransferase; D010450:Endopeptidases