PURIFICATION OF PHOSPHOLIPASE-C BY HYDROPHOBIC INTERACTION AFFINITY-CHROMATOGRAPHY

被引：5

作者：

CHAN, ECS ^{[1
]}

SIBOO, IR ^{[1
]}

SIBOO, R ^{[1
]}

机构：

[1] MCGILL UNIV, DEPT MICROBIOL & IMMUNOL, 3775 UNIV ST, MONTREAL H3A 2B4, QUEBEC, CANADA

来源：

JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS | 1991年 / 568卷 / 01期

基金：

英国医学研究理事会;

关键词：

D O I：

10.1016/0378-4347(91)80342-A

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A simple procedure is described for the purification of phosphatidylcholine-hydrolyzing phospholipase C(PLC). Lecithin, the substrate for PLC, was ligated hydrophobically to octyl-Sepharose in 2 M (NH4)2SO4. The washed lecithin-conjugated resin was then used to purify PLC from crude preparations by affinity chromatography. PLC binds to the lecithin moiety in the presence of Zn2+ and is eluted with an acidic buffer containing EDTA. PLC activity was recovered in the eluate. Both sodium dodecyl sulphate polyacrylamide gel electrophoresis and pI electrofocusing showed that the eluate contained a single monomeric protein with an apparent molecular mass of 66 kDa and a pI of 5.5.

引用

页码：85 / 92

页数：8

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