Assay of the pyruvate dehydrogenase complex by coupling with recombinant chicken liver arylamine N-acetyltransferase

Anal Biochem. 2006 Sep 1;356(1):44-50. doi: 10.1016/j.ab.2006.06.017. Epub 2006 Jun 30.

Abstract

The activity of the pyruvate dehydrogenase complex has long been determined in some laboratories by coupling the production of acetyl-coenzyme A (acetyl-CoA) to the acetylation of 4-aminoazobenzene-4'-sulfonic acid by arylamine N-acetyltransferase. The assay has some advantages, but its use has been limited by the need for large amounts of arylamine N-acetyltransferase. Here we report production of recombinant chicken liver arylamine N-acetyltransferase and optimization of its use in miniaturized assays for the pyruvate dehydrogenase complex and its kinase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arylamine N-Acetyltransferase / analysis
  • Arylamine N-Acetyltransferase / genetics
  • Base Sequence
  • Chickens
  • DNA / genetics
  • Liver / enzymology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Myocardium / enzymology
  • Protein Kinases / analysis
  • Protein-Serine-Threonine Kinases
  • Pyruvate Dehydrogenase (Acetyl-Transferring) Kinase
  • Pyruvate Dehydrogenase Complex / analysis*
  • Recombinant Proteins / analysis
  • Recombinant Proteins / genetics

Substances

  • Pyruvate Dehydrogenase (Acetyl-Transferring) Kinase
  • Pyruvate Dehydrogenase Complex
  • Recombinant Proteins
  • DNA
  • Arylamine N-Acetyltransferase
  • Protein Kinases
  • Protein-Serine-Threonine Kinases