Neskovic N M, Sarlieve L L, Mandel P
Biochim Biophys Acta. 1976 Apr 8;429(2):342-51. doi: 10.1016/0005-2744(76)90282-5.
A procedure for the purification of UDPgalactose--2-hydroxyacylsphingosine galactosyltransferase (EC 2.4.1.45) including detergent extraction, ion-excharge chromatography and proteolytic digestion was developed. The active fraction obtained by this procedure had about 100 times higher specific activity than microsomes. Enzymic activity resisted destruction by pronase treatment at 4 degrees C. Agarose gel chromatography indicated the presence of an enzyme-phospholipid-detergent complex with a molecular weight between 400 000 and 500 000. Intact phospholipids seemed to be required for full enzymic activity as evidenced by the drastic loss of activity upon treatment with phospholipase A or C.
已开发出一种纯化UDP-半乳糖-2-羟酰基鞘氨醇半乳糖基转移酶（EC 2.4.1.45）的方法，包括去污剂提取、离子交换色谱法和蛋白水解消化。通过该方法获得的活性级分比微粒体的比活性高约100倍。酶活性在4℃下经链霉蛋白酶处理后仍能抵抗破坏。琼脂糖凝胶色谱法表明存在一种分子量在400 000至500 000之间的酶-磷脂-去污剂复合物。完整的磷脂似乎是充分酶活性所必需的，这一点在用磷脂酶A或C处理后活性急剧丧失得到了证明。
