Characterization of the inhibitor sensitivity of the coenzyme A transport system in isolated rat heart mitochondria.
The effect of protein labeling agents on coenzyme A (CoA) transport into isolated rat heart mitochondria was studied. CoA transport was substantially inhibited by sulfhydryl reagents (mersalyl, pCMB) as well as by the tyrosine-selective reagent N-acetylimidazole. The effect of pCMB was reversed by DTT. Moreover, CoA uptake was completely abolished by agents selective for lysine and amino terminal residues (pyridoxal 5-phosphate, dansyl chloride). In contrast arginine-selective reagents (2, 3-butanedione, phenylglyoxal) caused considerably less inhibition of CoA uptake. Moreover, partial inhibition of transport was observed with the stilbene disulfonic acid derivatives DIDS and SITS. Finally, measurement of the effects of the labeling agents on the mitochondrial membrane potential indicated that the inhibition of CoA transport into mitochondria is not a secondary effect that arises from an alteration in the electric potential gradient across the inner mitochondrial membrane. These results provide the first information on the types of amino acid residues that may be essential to the CoA transport mechanism and provide additional support for the existence of a CoA transport protein within the mitochondrial inner membrane. Furthermore, the identification of effective inhibitors of the CoA transport system will greatly facilitate the functional reconstitution of this transporter in a proteoliposomal system following its solubilization and purification.
Tahiliani
A G
AG
Department of Biomedical Sciences, College of Allied Health, University of South Alabama, Mobile 36688.
Keene
T
T
Kaplan
R S
RS
D000818:Animals; D001120:Arginine / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D003065:Coenzyme A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism; D008929:Mitochondria, Heart / Q000201:enzymology*; D051381:Rats; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D013439:Sulfhydryl Reagents; D001120:Arginine; D008239:Lysine; D003065:Coenzyme A