title: Purification and characterization of the enzymes of bile acid conjugation from fish liver.
abstract: 1. The two steps in bile acid conjugation have been studied in subcellular fractions of liver from three species of fish; vermillion rockfish, canary rockfish and ling codfish. 2. The bile acid: coenzyme A (CoA) ligase activity in homogenates and isolated microsomes is undetectable due to indeterminate factors. 3. A purification scheme is presented which eliminates the interfering factors. The purified ligase was found to have a lower affinity for bile acids as compared to the mammalian form and to be present in much lower titer. 4. Since it appears to be the rate controlling enzyme in all species, it is expected that the rate of bile acid conjugation is much slower in non-mammalian liver as compared to mammalian liver. 5. The bile acid-CoA:taurine N-acyltransferase was found to exist as a dimer of molecular weight 100,000, in contrast to the monomeric mammalian forms. 6. The only major kinetic difference is that the fish liver forms have rates of glycine conjugation which are only 1-2% of the rate with taurine, in part due to a very high Km for glycine.
authors.affiliation: Liver Study Unit, Veterans Administration Medical Center, San Francisco, CA 94121.
authors.forename: D A
authors.initials: DA
authors.lastname: Vessey
authors.forename: A M
authors.initials: AM
authors.lastname: Benfatto
authors.forename: E
authors.initials: E
authors.lastname: Zerweck
authors.forename: C
authors.initials: C
authors.lastname: Vestweber
mesh_terms: D000217:Acyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001647:Bile Acids and Salts; D003066:Coenzyme A Ligases / Q000302:isolation & purification / Q000378:metabolism*; D005399:Fishes / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008322:Mammals; D008970:Molecular Weight; D013045:Species Specificity
chemical_list: D001647:Bile Acids and Salts; D000217:Acyltransferases; C025118:cholyl CoA glycine-taurine N-acyltransferase; D003066:Coenzyme A Ligases; C019890:choloyl-CoA synthetase