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

Research paper by Nam Ho NH Jeoung, Paresh C PC Sanghani, Lanmin L Zhai, Robert A RA Harris

Indexed on: 25 Jul '06Published on: 25 Jul '06Published in: Analytical Biochemistry: Methods in the Biological Sciences



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.