Characterization of the acyl-CoA:amino acid N-acyltransferases from primate liver mitochondria.
The acyl-CoA:amino acid N-acyl-transferases were partially purified from human liver mitochondria. The aralkyl transferase (ArAlk) had glycine conjugating activity toward the following compounds: benzoyl-CoA > butyryl-CoA, salicylyl-CoA > heptanoyl-CoA, indoleacetyl-CoA. Its kinetic properties and responses to salt were very similar to those of bovine ArAlk. Further, its molecular weight was found to be similar to that of the bovine enzyme, in contrast to reports from other laboratories. Thus, it was concluded that the human and bovine ArAlk are not significantly different. The human arylacetyl transferase (AAc) had glutamine conjugating activity toward phenylacetyl-CoA, but only 3-5% as much activity toward indoleacetyl-CoA or 1-naphtylacetyl-CoA, respectively. While this was similar to the bovine AAc, the two forms differed in several respects. First, the human liver AAc was insensitive to salts. Second, glycination of phenylacetyl-CoA by human AAc could only be detected at a high concentration of glycine (50 mM), and the rates were < 2% of the rate of glutamination. In contrast, glycine conjugation predominates with bovine AAc. Kinetic analysis of the glutamination of phenylacetyl-CoA by human AAc revealed a KD for phenylacetyl-CoA of 14 microM and a Km for glutamine of 120 mM. These values indicate that the human AAc is not more efficient at glutamination than the AAc from bovine liver. An AAc was purified from rhesus monkey liver and found to have similar kinetic constants to the human form. This indicates that nonprimate enzymes do not have a defect in glutamine conjugation.(ABSTRACT TRUNCATED AT 250 WORDS)
Kelley
M
M
Liver Study Unit, Veterans Affairs Medical Center, San Francisco, CA 94121.
Vessey
D A
DA
D000123:Acetyltransferases / Q000378:metabolism*; D000217:Acyltransferases / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D006801:Humans; D007525:Isoelectric Focusing; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008253:Macaca mulatta; D008930:Mitochondria, Liver / Q000201:enzymology*; D008970:Molecular Weight; D013045:Species Specificity; D013379:Substrate Specificity
D004228:Dithionitrobenzoic Acid; D000217:Acyltransferases; D000123:Acetyltransferases; C078176:aralkyl-CoA-glycine N-acyltransferase; C067287:arylacetyl-CoA N-acetyltransferase