A high-performance liquid chromatography method for the assay of cytidine monophosphate-sialic acid synthetase.
An HPLC method has been developed for the assay of cytidine monophosphate-sialic acid synthetase (EC 2.7.7.43) using ion-pair chromatography and gradient elution. This procedure permits the assay of alternative substrates and inhibitors of the enzyme and is not subject to the limitation of the colorimetric method. The newly synthesized N-acetyl-9-deoxy-9-fluoro-D-neuraminic acid was found to be a good substrate of the enzyme with a Km of 6.35 mM as compared to 1.84 mM for N-acetylneuraminic acid.
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003124:Colorimetry; D003568:Cytidine Monophosphate / Q000378:metabolism; D007700:Kinetics; D000216:N-Acylneuraminate Cytidylyltransferase / Q000032:analysis*; D009713:Nucleotidyltransferases / Q000032:analysis*; D012794:Sialic Acids / Q000378:metabolism
D012794:Sialic Acids; D009713:Nucleotidyltransferases; D000216:N-Acylneuraminate Cytidylyltransferase; D003568:Cytidine Monophosphate