PURIFICATION OF RAT-LIVER GLUCOKINASE

被引：2
作者：
TOYODA, Y
MIWA, I
KAMIYA, M
OGISO, S
OKUDA, J
机构：
[1] Department of Clinical Biochemistry, Faculty of Pharmacy, Meijo University, Nagoya
来源：
PREPARATIVE BIOCHEMISTRY | 1994年 / 24卷 / 3-4期
关键词：
D O I：
10.1080/10826069408010095
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
A new purification method for rat liver glucokinase was developed. Glucokinase was purified to homogeneity in a yield of 70% in 5 days. The procedure consists of DEAF-cellulose ion-exchange chromatography, QAE-Toyopearl ion-exchange chromatography, glucosamine-Sepharose affinity chromatography, and HiLoad Superdex 200 gel filtration. Purified glucokinase had a specific activity of 200 units/mg protein and was highly stable in the presence of 100 mM glucose, 300 mM KCl, and 20% glycerol. We found that some of the methionine residues of glucokinase were oxidized to methionine sulfoxide residues during dialysis in the presence of glucose. It would appear that this oxidation is caused by formation of hydroxyl radicals in the presence of glucose and contaminating transition metal(s).
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页码：225 / 236
页数：12
相关论文
共 19 条
[1]  
ANDREONE TL, 1989, J BIOL CHEM, V264, P363
[2]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[3]  
GROSSMAN SH, 1974, J BIOL CHEM, V249, P3055
[4]   METHIONINE OXIDATION AND INACTIVATION OF ALPHA-1-PROTEINASE INHIBITOR BY CU2+ AND GLUCOSE [J].
HALL, PK ;
ROBERTS, RC .
BIOCHIMICA ET BIOPHYSICA ACTA, 1992, 1121 (03) :325-330
[5]  
HARA H, 1986, CHEM PHARM BULL, V34, P4731
[6]   PURIFICATION IN HIGH-YIELD AND CHARACTERIZATION OF RAT HEPATIC GLUCOKINASE [J].
HOLROYDE, MJ ;
ALLEN, MB ;
STORER, AC ;
WARSY, AS ;
CHESHER, JME ;
TRAYER, IP ;
CORNISHBOWDEN, A ;
WALKER, DG .
BIOCHEMICAL JOURNAL, 1976, 153 (02) :363-+
[7]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+
[8]   GLUCOKINASE AS GLUCOSE SENSOR AND METABOLIC SIGNAL GENERATOR IN PANCREATIC BETA-CELLS AND HEPATOCYTES [J].
MATSCHINSKY, FM .
DIABETES, 1990, 39 (06) :647-652
[9]   NEW PERSPECTIVES ON PANCREATIC-ISLET GLUCOKINASE [J].
MEGLASSON, MD ;
MATSCHINSKY, FM .
AMERICAN JOURNAL OF PHYSIOLOGY, 1984, 246 (01) :E1-E13
[10]   HIGH-YIELD PURIFICATION OF GLUCOKINASE FROM RAT-LIVER [J].
MIWA, I ;
MITSUYAMA, S ;
TOYODA, Y ;
MURATA, T ;
OKUDA, J .
PREPARATIVE BIOCHEMISTRY, 1990, 20 (02) :163-178
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