Mori M, Tatibana M
J Biochem. 1975 Jul;78(1):239-42.
Glutamine-dependent carbamyl phosphate synthetase [EC 2.7.2.9] was purified 1,300-fold from rat ascites hepatoma cells (AH 13) as a multienzyme complex with aspartate transcarbamylase[EC 2.1.3.2] and dihydroorotase[EC 3.5.2.3], using dimethyl sulfoxide, glycerol, and dithiothreitol as stabilizers. The purified complex was essentially homogeneous on agarose-acrylamide composite gel electrophoresis and analytical ultracentrifugation. Its molecular weight was estimated to be about 870,000 by sedimentation equilibrium studies. After alkylation with iodoacetamide or reduction with 0.6% dithiothreitol at 100 degrees, the complex gave a single band on polyacrylamide gel electrophoresis in sodium dodecyl sulfate in a position corresponding to a molecular weight of 210,000. These results indicate that the complex consists of four subunits of similar size.
谷氨酰胺依赖性氨甲酰磷酸合成酶[EC 2.7.2.9]从大鼠腹水肝癌细胞（AH 13）中作为与天冬氨酸转氨甲酰酶[EC 2.1.3.2]和二氢乳清酸酶[EC 3.5.2.3]的多酶复合物纯化了1300倍，使用二甲基亚砜、甘油和二硫苏糖醇作为稳定剂。纯化后的复合物在琼脂糖-丙烯酰胺复合凝胶电泳和分析超速离心中基本呈均一状态。通过沉降平衡研究估计其分子量约为870,000。用碘乙酰胺烷基化或在100℃下用0.6%二硫苏糖醇还原后，该复合物在十二烷基硫酸钠聚丙烯酰胺凝胶电泳上呈现一条带，其位置对应于分子量为210,000。这些结果表明该复合物由四个大小相似的亚基组成。
