Choy P C, Farren S B, Vance D E
Can J Biochem. 1979 Jun;57(6):605-12. doi: 10.1139/o79-076.
Two forms of CTP:phosphocholine cytidylyltransferase were identified in rat liver cytosol by gel filtration chromatography. The low molecular weight form (L form) is the major form in fresh cytosol. The enzyme associates into a high molecular weight form (H form) upon storage of the cytosol at 4 degrees C. Aggregation of the purified L form of cytidylyltransferase is caused by total rat liver lipids, neutral lipids, diacylglycerol, or phosphatidylglycerol. Diacylglycerol was the only lipid isolated from the rat liver that caused aggregation of the purified enzyme. Although the addition of diacylglycerol to the cytosol did not change the amount of aggregation of the enzyme, a 2.5-fold increase in H form was observed in cytosol pretreated with phospholipase C, or in cytosol from rats fed a high cholesterol diet. In both of these cytosolic preparations, the concentration of diacylglycerol was elevated twofold. Phosphatidylglycerol did not seem to affect the association of the enzyme in cytosol since it is present in very low concentrations in the rat liver cytosol, and its degradation in cytosol by a specific phospholipase did not affect the rate of aggregation. The results suggest that diacylglycerol in an appropriate form is required for association of cytidylyltransferase in rat liver cytosol.
通过凝胶过滤色谱法在大鼠肝脏胞质溶胶中鉴定出两种形式的CTP:磷酸胆碱胞苷转移酶。低分子量形式(L型)是新鲜胞质溶胶中的主要形式。当胞质溶胶在4℃储存时,该酶会结合形成高分子量形式(H型)。胞苷转移酶纯化的L型的聚集是由全大鼠肝脏脂质、中性脂质、二酰基甘油或磷脂酰甘油引起的。二酰基甘油是从大鼠肝脏中分离出的唯一能引起纯化酶聚集的脂质。虽然向胞质溶胶中添加二酰基甘油并没有改变酶的聚集量,但在用磷脂酶C预处理的胞质溶胶或喂食高胆固醇饮食的大鼠的胞质溶胶中,观察到H型增加了2.5倍。在这两种胞质溶胶制剂中,二酰基甘油的浓度都升高了两倍。磷脂酰甘油似乎不影响该酶在胞质溶胶中的结合,因为它在大鼠肝脏胞质溶胶中的浓度非常低,并且其在胞质溶胶中被特异性磷脂酶降解并不影响聚集速率。结果表明,大鼠肝脏胞质溶胶中胞苷转移酶的结合需要适当形式的二酰基甘油。