title: A new assay for L-asparagine synthetase.
abstract: A fast, relatively inexpensive method of measuring the enzymatic formation of L-asparagine from L-aspartate is presented. This radiochemical assay requires simple manipulations making it ideal for working with large numbers of samples, while maintaining high sensitivity and reproducibility. A mechanism similar to the enzymatic beta-decarboxylation of aspartate is utilized but in a nonenzymatic reaction. In the presence of pyridoxal and A13+ ions, the 14C of L-[4-14C]aspartate is decarboxylated while L-[4-14C]asparagine remains intact. This assay is shown to be suitable for measuring mammalian L-asparagine synthetase activity, while not requiring the isolation of assay enzymes.
mesh_terms: D000535:Aluminum; D001216:Asparagine; D001217:Aspartate-Ammonia Ligase / Q000032:analysis*; D001224:Aspartic Acid; D002250:Carbon Radioisotopes; D003653:Decarboxylation; D008025:Ligases / Q000032:analysis*; D008722:Methods; D011730:Pyridoxal
chemical_list: D002250:Carbon Radioisotopes; D001224:Aspartic Acid; D011730:Pyridoxal; D001216:Asparagine; D000535:Aluminum; D008025:Ligases; D001217:Aspartate-Ammonia Ligase