Development of a radiolabeled ATP assay for carboxylic acid:CoA ligases and its use in the characterization of the xenobiotic carboxylic acid:CoA ligases of bovine liver mitochondria.


Journal Article

A radiolabeled ATP assay was developed for measuring carboxylic acid:CoA ligase activity. The assay was designed to measure the formation of [gamma-33P]pyrophosphate from [gamma-33P]ATP in the course of the reaction. The assay was linear with protein concentration, and rates as low as 1 pmol/min were measurable. Rates determined with this assay were in agreement with rates determined with [14C]carboxylic acids. The assay was used to characterize the substrate specificity of the XL-I, XL-II, and XL-III ligases from bovine liver mitochondria. Forty carboxylic acids were tested for activity. The enzymes differed in their substrate specificities with XL-I and XL-II being the most similar and XL-III having the broadest specificity. This study has uncovered 19 new carboxylic acids that are substrates for these enzymes.

Full Text

Cited Authors

  • Vessey, DA; Kelley, M; Lau, E

Published Date

  • January 1998

Published In

Volume / Issue

  • 12 / 3

Start / End Page

  • 151 - 155

PubMed ID

  • 9522274

Pubmed Central ID

  • 9522274

Electronic International Standard Serial Number (EISSN)

  • 1099-0461

International Standard Serial Number (ISSN)

  • 1095-6670

Digital Object Identifier (DOI)

  • 10.1002/(sici)1099-0461(1998)12:3<151::aid-jbt3>;2-k


  • eng