Analytical Biochemistry 2007-09-01

Dephospho-CoA kinase provides a rapid and sensitive radiochemical assay for coenzyme A and its thioesters.

Caryn Wadler, John E Cronan

Index: Anal. Biochem. 368 , 17-23, (2007)

Full Text: HTML

Abstract

A new approach to determine in vivo pools of coenzyme A (CoA) and short chain acyl-CoA thioesters is reported. The metabolites released by extraction with trichloroacetic acid are recovered and quantitatively dephosphorylated by treatment with shrimp alkaline phosphatase. Following phosphatase removal, the dephosphorylated CoA metabolites are quantitatively rephosphorylated by treatment with [gamma-33P]ATP plus a dephospho-CoA kinase. The resulting radioactive CoA metabolites are then separated by reverse-phase high-performance liquid chromatography and quantitated by scintillation counting. Due to the enzymatic radiophosphorylation, the assay is specific for CoA and its short chain thioesters and is sensitive to sub-picomole levels of these compounds.

Related Compounds

  • 3'-O-dephosphono-c...
  • SODIUM AMMON...

Related Articles:

Simultaneous high-performance liquid chromatography determination of coenzyme A, dephospho-coenzyme A, and acetyl-coenzyme A in normal and pantothenic acid-deficient rats.

2012-11-15

[Anal. Biochem. 430(2) , 151-5, (2012)]

Identification of the active site of phosphoribosyl-dephospho-coenzyme A transferase and relationship of the enzyme to an ancient class of nucleotidyltransferases.

2000-10-31

[Biochemistry 39(43) , 13233-40, (2000)]

Purification and characterization of phosphopantetheine adenylyltransferase from Escherichia coli.

1999-09-17

[J. Biol. Chem. 274(38) , 27105-11, (1999)]

Hydrolysis and absorption of pantothenate and its coenzymes in the rat small intestine.

1983-10-01

[J. Nutr. 113(10) , 2107-15, (1983)]

Coenzyme A affects firefly luciferase luminescence because it acts as a substrate and not as an allosteric effector.

2005-10-01

[FEBS J. 272(20) , 5206-16, (2005)]

More Articles...