Purification and properties of carnitine acetyltransferases from bovine spermatozoa and heart.
To investigate the physical and kinetic properties of sperm carnitine acetyltransferase, the enzyme was purified from bovine spermatozoa and heart muscle. Carnitine acetyltransferase was purified 580-fold from ejaculated bovine spermatozoa to a specific activity of 85 units/mg protein (95% homogeneity). Sperm carnitine acetyltransferase was characterized as a single polypeptide of Mr 62,000 and pI 8.2. Heart carnitine acetyltransferase was purified 650-fold by the same procedure to a final specific activity of 71 units/mg protein. The kinetic properties of purified bovine sperm carnitine acetyltransferase were consistent with the proposed function of this enzyme in acetylcarnitine pool formation. Product inhibition by either acetyl-L-carnitine or CoASH was not sufficient to predict significant in vivo inhibition of acetyl transfer. At high concentrations of L-carnitine, bovine sperm and heart carnitine acetyltransferases were most active with propionyl- and butyryl-CoA substrates, although octanoyl-, iso-butyryl-, and iso-valeryl-CoA were acceptable substrates. Binding of one substrate was enhanced by the presence of the second substrate. Carnitine analogs that have significance in reproduction, such as phosphorylcholine and taurine, did not inhibit carnitine acetyltransferase. Bovine sperm and heart carnitine acetyltransferases were indistinguishable on the basis of purification behavior, pI, pH optima, kinetic properties, acyl-CoA specificity, and sensitivity to sulfhydryl reagents and divalent cations; thus there was no indication that bovine sperm carnitine acetyltransferase is a sperm-specific isozyme.
D000123:Acetyltransferases / Q000302:isolation & purification*; D000818:Animals; D002331:Carnitine / Q000494:pharmacology; D002332:Carnitine O-Acetyltransferase / Q000302:isolation & purification* / Q000378:metabolism; D002413:Cations, Divalent; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D007536:Isomerism; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology*; D009928:Organ Specificity; D013094:Spermatozoa / Q000201:enzymology*
D002413:Cations, Divalent; D000123:Acetyltransferases; D002332:Carnitine O-Acetyltransferase; D002331:Carnitine