A carnitine/acylcarnitine translocase assay applicable to biopsied muscle specimens without requiring mitochondrial isolation.
A simple method for assaying the mitochondrial carnitine/acylcarnitine translocase of muscles that needs only few milligrams of fresh tissue is described. The procedure involves monitoring of the sulphobetaine (an inhibitor of the translocase)-sensitive acetylation of sub-saturating concentrations of carnitine in the medium, linked to the oxidation of [2-14C]pyruvate in the presence of malonate. Conditions affecting the reliability of the outlined procedure and the ancillary information to be collected, namely the activities of pyruvate oxidase system and carnitine acetyltransferase, for detecting possible deficiency of the translocase are described, together with data on the translocase activity in human skeletal muscle, in rat red and white skeletal muscles and in rat heart. The concepts outlined should allow development of assays of other mitochondrial transporters that also would require neither isolation of mitochondria nor availability of a large quantity of tissue, both of which are otherwise needed at present.
D000107:Acetylation; D000108:Acetylcarnitine / Q000378:metabolism; D000328:Adult; D000368:Aged; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D001692:Biological Transport; D002331:Carnitine / Q000378:metabolism; D002333:Carnitine Acyltransferases; D006801:Humans; D008297:Male; D008875:Middle Aged; D008931:Mitochondria, Muscle / Q000201:enzymology; D009124:Muscle Proteins / Q000378:metabolism; D009132:Muscles / Q000201:enzymology*; D009206:Myocardium / Q000201:enzymology; D011773:Pyruvates / Q000378:metabolism; D019289:Pyruvic Acid; D051381:Rats; D014166:Transferases / Q000378:metabolism*
D009124:Muscle Proteins; D011773:Pyruvates; D000108:Acetylcarnitine; D019289:Pyruvic Acid; D014166:Transferases; D002333:Carnitine Acyltransferases; D002331:Carnitine