Zhao M, Rua D, Hajra A K, Greenberg M L
Department of Biological Sciences, Wayne State University, Detroit, Michigan 48202, USA.
Anal Biochem. 1998 Apr 10;258(1):48-52. doi: 10.1006/abio.1997.2559.
Cytidine 5'-diphospho-1,2-diacyl-sn-glycerol (CDP-diacylglycerol; CDP-DG) is an important intermediate in the biosynthesis of the major glycerophosphate-based phospholipids of prokaryotes and eukaryotes. This compound is expensive to purchase and inefficient to prepare chemically. Radiolabeled CDP-diacylglycerol is unavailable commercially. We describe a simple and inexpensive method to synthesize [3H]CDP-DG enzymatically. The three-step enzymatic procedure includes phosphorylation of [3H]glycerol to sn-[3H]glycerol 3-phosphate (G3P) by glycerokinase,acylation of [3H]G3P to [3H]phosphatidic acid (PA) by G3P acyltransferase, and conversion of [3H]PA and CTP to [3H]CDP-DG by CDP-DG synthase. This procedure is considerably less labor intensive and less expensive than is chemical synthesis, and the yield is at least 30%.
胞苷5'-二磷酸-1,2-二酰基-sn-甘油（CDP-二酰甘油；CDP-DG）是原核生物和真核生物中主要基于甘油磷酸的磷脂生物合成的重要中间体。该化合物购买成本高昂，化学合成效率低下。放射性标记的CDP-二酰甘油无法从商业渠道获得。我们描述了一种简单且廉价的酶促合成[3H]CDP-DG的方法。这个三步酶促过程包括通过甘油激酶将[3H]甘油磷酸化为sn-[3H]甘油3-磷酸（G3P），通过G3P酰基转移酶将[3H]G3P酰化为[3H]磷脂酸（PA），以及通过CDP-DG合酶将[3H]PA和CTP转化为[3H]CDP-DG。该过程相比化学合成，劳动强度大大降低且成本更低，产率至少为30%。
