A coupled assay for bile acid:CoA ligase.
A new assay for the enzyme bile acid:CoA ligase is presented. The new assay is designed to supplant the existing radiometric assays which require radiolabeled bile acids. The new assay couples the formation of bile acid-CoA to its glycination in a reaction catalyzed by bile acid-CoA:glycine N-acyltransferase. The coupling reaction utilizes [14C]glycine and the bile acid-CoA is quantitatively converted to [14C]glycobile acid. The [14C]glycobile acid is isolated by solvent extraction and quantitated by liquid scintillation counting. The method is shown to be accurate, highly sensitive, and applicable to a wide variety of bile acids.
Vessey
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DA
Liver Study Unit, Veterans Affairs Medical Center, San Francisco, California 94121.
Erdman
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A
Kelley
M
M
St Claire
T
T
D000217:Acyltransferases / Q000378:metabolism; D000818:Animals; D002417:Cattle; D003066:Coenzyme A Ligases / Q000032:analysis*; D006168:Guinea Pigs; D008862:Microsomes, Liver / Q000201:enzymology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000217:Acyltransferases; C025118:cholyl CoA glycine-taurine N-acyltransferase; D003066:Coenzyme A Ligases; C070656:bile acid-CoA ligase; C019890:choloyl-CoA synthetase