A high-performance liquid chromatography assay for asparagine synthetase.
A highly sensitive method for assaying asparagine synthetase and its glutaminase activity is presented. The amino acids L-asparagine, L-aspartate, L-glutamate, and L-glutamine, are separated by derivatization with o-phthaldialdehyde followed by reversed-phase high-performance liquid chromatography on an Altex ultrasphere-ODS C18 column. The elution is isocratic and the mobile phase used is 50 mM sodium acetate buffer (pH 5.9) with 30% methanol. This assay can easily detect picomoles of asparagine, which may be difficult to do with the other assays that have been described.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001216:Asparagine / Q000032:analysis; D001217:Aspartate-Ammonia Ligase / Q000032:analysis*; D001224:Aspartic Acid / Q000032:analysis; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005971:Glutamates / Q000032:analysis; D005973:Glutamine / Q000032:analysis; D008025:Ligases / Q000032:analysis*; D010179:Pancreas / Q000201:enzymology; D011730:Pyridoxal / Q000032:analysis; D009764:o-Phthalaldehyde
D000596:Amino Acids; D005971:Glutamates; D005973:Glutamine; D001224:Aspartic Acid; D011730:Pyridoxal; D009764:o-Phthalaldehyde; D001216:Asparagine; D008025:Ligases; D001217:Aspartate-Ammonia Ligase