Gatt S, Dinur T, Barenholz Y
Biochim Biophys Acta. 1978 Sep 28;530(3):503-7. doi: 10.1016/0005-2760(78)90169-8.
A colored derivative of sphingomyelin was synthesized and used as substrate for several sphingomyelinases. The compound is N-omega-trinitrophenyl-aminolaurylsphingosylphosphorylcholine. The rate of hydrolysis of this substrate was compared to that of bovine brain sphingomyelin, labelled with tritium in the choline moiety. The following enzyme preparations were used: homogenate-less debris of brain, assayed at pH 5.0 or 7.4; a solubilized preparation derived from rat brain lysosomes, assayed at pH 5.0 and a purified enzyme of Staphylococcus aureus. With all preparations, the rates of hydrolysis of the yellow derivative were very similar to those of the brain sphingomyelin. Extracts of skin fibroblasts of normal and Niemann-Pick patients as well as amniotic cells were also used. Again, the rates of hydrolysis of the yellow derivative practically equalled those using brain sphingomyelin.
合成了一种鞘磷脂的有色衍生物，并将其用作几种鞘磷脂酶的底物。该化合物为N-ω-三硝基苯基-氨基月桂酰鞘氨醇磷酸胆碱。将该底物的水解速率与在胆碱部分用氚标记的牛脑鞘磷脂的水解速率进行了比较。使用了以下酶制剂：脑的无匀浆残渣，在pH 5.0或7.4下进行测定；源自大鼠脑溶酶体的可溶制剂，在pH 5.0下进行测定；以及金黄色葡萄球菌的纯化酶。对于所有制剂，黄色衍生物的水解速率与脑鞘磷脂的水解速率非常相似。还使用了正常人和尼曼-匹克病患者的皮肤成纤维细胞提取物以及羊膜细胞。同样，黄色衍生物的水解速率实际上与使用脑鞘磷脂时的水解速率相等。
