Cox J W, Snyder W R, Horrocks L A
Chem Phys Lipids. 1979 Dec;25(4):369-80. doi: 10.1016/0009-3084(79)90075-6.
Spectrophotometric assays of esterases are sensitive, rapid, and quite specific when thioester substrates are used. Glycerophospholipids with thiophosphoester bonds may be useful as substrates for phospholipase C (EC 3.1.4.3). These have been made from mercaptoglycerol and mercaptoethanol. The thiols were oxidized to disulfides, acylated, and reduced with dithiothreitol. Phosphocholine derivatives were made by the classical methods for oxyphosphoesters. The phosphatidyl choline analogue was converted to the phosphatidyl ethanolamine analogue by transphosphatidylation with cabbage phospholipase D and ethanolamine. Structures were proved with enzymic hydrolysis, infrared spectra, TLC behavior, and elemental analyses. The synthesized compounds were rac-1-S-phosphocholine-2,3-O-didecanoyl-1-mercapto-2,3-propanediol, 1-S-phosphoethanolamine-2,3-O-didecanoyl-1-mercapto-2,3-propanediol, and 1-S-phosphocholine-2-O-hexadecanoyl-1-mercapto-2-ethanol.
当使用硫酯底物时，酯酶的分光光度测定法灵敏、快速且具有相当的特异性。带有硫代磷酸酯键的甘油磷脂可能用作磷脂酶C（EC 3.1.4.3）的底物。这些已由巯基甘油和巯基乙醇制备而成。硫醇被氧化成二硫化物，进行酰化反应，并用二硫苏糖醇还原。磷酸胆碱衍生物通过经典的氧代磷酸酯方法制备。通过用卷心菜磷脂酶D和乙醇胺进行转磷脂酰化反应，将磷脂酰胆碱类似物转化为磷脂酰乙醇胺类似物。通过酶促水解、红外光谱、薄层色谱行为和元素分析来证实结构。合成的化合物为rac-1-S-磷酸胆碱-2,3-O-二癸酰基-1-巯基-2,3-丙二醇、1-S-磷酸乙醇胺-2,3-O-二癸酰基-1-巯基-2,3-丙二醇和1-S-磷酸胆碱-2-O-十六酰基-1-巯基-2-乙醇。
