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[14C]磷脂酰胆碱在脂质体与人血浆高密度脂蛋白之间的转移。利用快速转移测定法对一种转移刺激血浆因子进行部分纯化。

Transfer of [14C]phosphatidylcholine between liposomes and human plasma high density lipoprotein. Partial purification of a transfer-stimulating plasma factor using a rapid transfer assay.

作者信息

Damen J, Regts J, Scherphof G

出版信息

Biochim Biophys Acta. 1982 Sep 14;712(3):444-52. doi: 10.1016/0005-2760(82)90271-5.

Abstract

A simple method was developed for the rapid determination of [14C]phosphatidylcholine transfer from small unilamellar liposomes to human plasma HDL, based on the selective precipitation of liposomes by heparin and MnCl2. The assay was utilized to monitor the progress in the partial purification of a phospholipid transfer factor from human plasma. The purification procedure included ultracentrifugation at d = 1.25 g/ml, hydrophobic chromatography on phenyl-Sepharose, affinity chromatography on heparin-Sepharose and gel filtration. The partially purified protein(s) catalyzed the net transfer of phospholipid from small unilamellar phosphatidylcholine liposomes to isolated HDL. The transfer of [14C]phosphatidylcholine from liposomes consisting of phosphatidylcholine/phosphatidylserine/cholesterol (molar ratio, 4:1:5) to HDL was stimulated without affecting the permeability barrier of the liposomal membranes and is, therefore, taken to represent exchange with HDL phospholipid rather than net transfer.

摘要

基于肝素和氯化锰对脂质体的选择性沉淀作用,开发了一种简单的方法,用于快速测定[¹⁴C]磷脂酰胆碱从小单层脂质体向人血浆高密度脂蛋白(HDL)的转移。该测定法用于监测从人血浆中部分纯化磷脂转移因子的进展情况。纯化过程包括在密度为1.25 g/ml下超速离心、在苯基琼脂糖上进行疏水色谱、在肝素琼脂糖上进行亲和色谱以及凝胶过滤。部分纯化的蛋白质催化了磷脂从小单层磷脂酰胆碱脂质体向分离的HDL的净转移。[¹⁴C]磷脂酰胆碱从由磷脂酰胆碱/磷脂酰丝氨酸/胆固醇(摩尔比为4:1:5)组成的脂质体向HDL的转移受到刺激,且不影响脂质体膜的通透性屏障,因此被认为代表与HDL磷脂的交换而非净转移。

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