Purification of Aspartate Transcabamylase from Drosophila melanogaster

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Abstract
A purification procedure is described by which aspartate transcarbamylase [EC 2.1.3.2] was obtained from cultured cells of D. melanogaster as part of a high MW enzyme complex. The complex contained several polypeptides. An antiserum directed against the complex enzyme inhibited in vitro the activity of aspartate transcarbamylase, carbamylphosphate synthetase [EC 2.7.2.9] and dihydro-orotase [EC 3.5.2.3], which co-purified on a sucrose gradient and by gel electrophoresis. A fast preparation procedure using this antiserum yielded a 220,000 MW protein in addition to the polypeptides present in the complex. A purification procedure is described to obtain aspartate transcarbamylase from 2nd instar Drosophila larvae. At this stage, the enzyme was not complexed with carbamylphosphate synthetase and dihydro-orotase but exhibited the same MW as the asparatate transcarbamylase moiety found in the high MW complex of cultured cells.