title: A capillary electrophoretic assay for acetyl coenzyme A carboxylase.
abstract: A simple off-column capillary electrophoretic (CE) assay for measuring acetyl coenzyme A carboxylase holoenzyme (holo-ACC) activity and inhibition was developed. The two reactions catalyzed by the holo-ACC components, biotin carboxylase (BC) and carboxyltransferase (CT), were simultaneously monitored in this assay. Acetyl coenzyme A (CoA), malonyl-CoA, adenosine triphosphate (ATP), and adenosine diphosphate (ADP) were separated by capillary electrophoresis, and the depletion of ATP and acetyl-CoA as well as the production of ADP and malonyl-CoA were monitored. Inhibition of holo-ACC by the BC inhibitor, 2-amino-N,N-dibenzyloxazole-5-carboxamide, and the carboxyltransferase inhibitor, andrimid, was confirmed using this assay. A previously reported off-column CE assay for only the CT component of ACC was optimized, and an off-column CE assay for the BC component of ACC also was developed.
authors: Department of Chemistry, Louisiana State University, Baton Rouge, LA 70803, USA.
authors: Sherrisse K
authors: SK
authors: Bryant
authors: Grover L
authors: GL
authors: Waldrop
authors: S Douglass
authors: SD
authors: Gilman
mesh_terms: D000105:Acetyl Coenzyme A / Q000378:metabolism; D000103:Acetyl-CoA Carboxylase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D055162:Biocatalysis; D019731:Carbon-Nitrogen Ligases / Q000378:metabolism; D019878:Carboxyl and Carbamoyl Transferases / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology; D020035:Holoenzymes / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D020035:Holoenzymes; D000105:Acetyl Coenzyme A; D000255:Adenosine Triphosphate; D019878:Carboxyl and Carbamoyl Transferases; D019731:Carbon-Nitrogen Ligases; C019791:biotin carboxylase; D000103:Acetyl-CoA Carboxylase