A PURIFICATION STRATEGY FOR INOSITOL 1,4,5-TRISPHOSPHATE 3-KINASE FROM RAT-LIVER BASED UPON HEPARIN INTERACTION CHROMATOGRAPHY

被引：12

作者：

CONIGRAVE, A

PATWARDHAN, A

BROOMHEAD, L

ROUFOGALIS, B

机构：

[1] UNIV SYDNEY,DEPT BIOCHEM,SYDNEY,NSW 2006,AUSTRALIA

[2] UNIV SYDNEY,DEPT PHYSIOL,SYDNEY,NSW 2006,AUSTRALIA

来源：

CELLULAR SIGNALLING | 1992年 / 4卷 / 03期

关键词：

INS (1,3,4,5)P(4); INS (1,4,5)P(3) 3-KINASE; CALMODULIN; HEPARIN; ENZYME PURIFICATION; RAT LIVER; PYROPHOSPHATE;

D O I：

10.1016/0898-6568(92)90070-O

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Rat liver inositol 1,4,5-trisphosphate [Ins (1,4,5)P3] 3-kinase was purified in high yield by a three-step procedure reliant upon chromatography on heparin and calmodulin agarose. Purified enzyme was' stable in the presence of the detergent 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulphonate (CHAPS) (0.1-0.5%) and the sulphhydryl reducing reagent dithiothreitol (DTT). The purified enzyme was activated 2-3-fold by Ca2+ (1-mu-M) in the presence of calmodulin. Pyrophosphate and heparin were identified as inhibitors of the enzyme.

引用

页码：303 / &

共 16 条

[1] GTP-GAMMA-S-STIMULATED HYDROLYSIS OF PHOSPHATIDYINOSITOL-4,5-BISPHOSPHATE BY SOLUBLE PHOSPHOLIPASE-C FROM HUMAN-PLATELETS REQUIRES SOLUBLE GTP-BINDING PROTEIN [J].