A method for the purification of milligram quantities of stable human phosphatidylcholine-cholesterol acyltransferase.
A method for the purification of milligram quantities of stable human phosphatidylcholine-cholesterol acyltransferase.
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一种纯化毫克量的稳定人磷脂酰胆碱-胆固醇酰基转移酶的方法。
DOI：
10.1042/bj1550583
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发表时间：
1976
期刊：
影响因子：
--
通讯作者：
L. Soloff
中科院分区：
文献类型：
--
作者：
K. Varma;L. Soloff
描述了一种处理3升人血浆以纯化磷脂酰胆碱-胆固醇酰基转移酶的方法。该方法包括 (NH4)2SO4 分级分离、柠檬酸处理以及 DEAE-纤维素和羟基磷灰石色谱法。在此阶段，酶制剂被纯化约。 8000倍。根据抗人血清免疫电泳测定，该制剂似乎不含脂蛋白，并且根据免疫扩散测定，白蛋白污染极少（少于 30 微克/毫克酶蛋白）。酶活性4天稳定，20天后大部分活性丧失。在5%聚丙烯酰胺凝胶上电泳，观察到一条有酶活性的快速移动条带和一条无酶活性的慢速移动条带。还存在微弱的白蛋白带。从 10 块凝胶上切下酶活性条带的提取物，然后合并并用 0.15 M-NaC1/4mM-磷酸钠（pH 7.0）提取，在 5% 聚丙烯酰胺凝胶上重新电泳时显示单条带。
A method for processing 3 litres of human plasma for the purification of phosphatidylcholine-cholesterol acyltransferase is described. The method involves (NH4)2SO4 fractionation, citric acid treatment, and DEAE-cellulose and hydroxyapatite chromatography. At this stage the enzyme preparation is purified approx. 8000-fold. This preparation appears to be free of lipoproteins as determined by immunoelectrophoresis against anti-human serum and is minimally contaminated with albumin (less than 30 mug/mg of enzyme protein) as determined by immunodiffusion. The activity of the enzyme was stable for 4 days, but most of its activity was lost after 20 days On electrophoresis on 5% polyacrylamide gel, a fast-moving band with enzyme activity and a slow-moving band with no enzyme activity was observed. A faint band of albumin was also present. Extracts of enzymically active bands cut from ten gels and then pooled and extracted with 0.15 M-NaC1/4mM-sodium phosphate, pH 7.0, showed a single band on re-electrophoresis on 5% polyacrylamide gel.