PURIFICATION AND CRYSTALLIZATION OF YEAST HEXOKINASE ISOENZYMES - CHARACTERIZATION OF DIFFERENT FORMS BY CHROMATOFOCUSING

被引：11

作者：

JACOB, L ^{[1
]}

BEECKEN, V ^{[1
]}

BARTUNIK, LJ ^{[1
]}

ROSE, M ^{[1
]}

BARTUNIK, HD ^{[1
]}

机构：

[1] UNIV FRANKFURT,INST MIKROBIOL,W-6000 FRANKFURT,GERMANY

来源：

JOURNAL OF CHROMATOGRAPHY | 1991年 / 587卷 / 01期

关键词：

D O I：

10.1016/0021-9673(91)85201-P

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

The yeast hexokinase isoenzymes PI and PII have been purified in large amounts (20 mg) from overproducing yeast strains. The purification procedures of hexokinase PI and PII include anion-exchange chromatography on DEAE-Sephacel and chromatofocusing on PBE 94, hydrophobic interaction chromatography on phenyl-Sepharose (necessary for the isolation of the isoenzyme PI); in the final step either a Mono Q HR 5/5 or a Fractogel EMD TMAE 650(S) column was used. Hexokinase preparations were characterized before cyrstallization by chromatofocusing on a Mono P HR 5/20 FPLC column, where different forms of hexokinase can be rapidly distinguished by their elution behaviour. From both purified hexokinase PI and PII, large crystals were grown that diffract X-rays to high resolution.

引用

页码：85 / 92

页数：8

共 26 条

[1] SEQUENCING A PROTEIN BY X-RAY CRYSTALLOGRAPHY .2. REFINEMENT OF YEAST HEXOKINASE-B COORDINATES AND SEQUENCE AT 2.1-A RESOLUTION [J].