title: Assay of the pyruvate dehydrogenase complex by coupling with recombinant chicken liver arylamine N-acetyltransferase.
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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
authors.forename: Nam Ho
authors.initials: NH
authors.lastname: Jeoung
authors.forename: Paresh C
authors.initials: PC
authors.lastname: Sanghani
authors.forename: Lanmin
authors.initials: L
authors.lastname: Zhai
authors.forename: Robert A
authors.initials: RA
authors.lastname: Harris
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001191:Arylamine N-Acetyltransferase / Q000032:analysis / Q000235:genetics; D001483:Base Sequence; D002645:Chickens; D004247:DNA / Q000235:genetics; D008099:Liver / Q000201:enzymology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009206:Myocardium / Q000201:enzymology; D011494:Protein Kinases / Q000032:analysis; D017346:Protein Serine-Threonine Kinases; D000081382:Pyruvate Dehydrogenase Acetyl-Transferring Kinase; D011768:Pyruvate Dehydrogenase Complex / Q000032:analysis*; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics
chemical_list: D000081382:Pyruvate Dehydrogenase Acetyl-Transferring Kinase; D011768:Pyruvate Dehydrogenase Complex; D011994:Recombinant Proteins; D004247:DNA; D001191:Arylamine N-Acetyltransferase; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases