Molecular cloning, heterologous expression, and characterization of human glyoxalase II

被引:68
作者
Ridderstrom, M
Saccucci, F
Hellmann, U
Bergman, T
Principato, G
Mannervik, B
机构
[1] UNIV UPPSALA,CTR BIOMED,DEPT BIOCHEM,S-75123 UPPSALA,SWEDEN
[2] UNIV ANCONA,FAC MED & CHIRURG,IST BIOL & GENET,I-60100 ANCONA,ITALY
[3] BIOMED CTR,LUDWIG INST CANC RES,S-75124 UPPSALA,SWEDEN
[4] KAROLINSKA INST,DEPT MED BIOCHEM & BIOPHYS,S-17177 STOCKHOLM,SWEDEN
关键词
D O I
10.1074/jbc.271.1.319
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 clone encoding glyoxalase II has been isolated from a human adult liver cDNA library, The sequence of 1011 base pairs consists of a full-length coding region of 780 base pairs, corresponding to a protein with a calculated molecular mass of 28,861 daltons. Identities (50-60%) were found to partial 5' and 3' cDNA sequences from Arabidopsis thaliana as well as within a limited region of glutathione transferase I cDNA from corn, A vector was constructed for heterologous expression of glyoxalase II in Escherichia coli. For optimal yield of enzyme, silent random mutations were introduced in the 5' coding region of the cDNA. A yield of 25 mg of glyoxalase II per liter of culture medium was obtained after affinity purification with immobilized glutathione. The recombinant enzyme had full catalytic activity and kinetic parameters indistinguishable from those of the native enzyme purified from human erythrocytes.
引用
收藏
页码:319 / 323
页数:5
相关论文
共 33 条
  • [1] PURIFICATION AND CHARACTERIZATION OF GLYOXALASE-II FROM HUMAN RED-BLOOD-CELLS
    ALLEN, RE
    THEODORE, WCL
    THORNALLEY, PJ
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1993, 213 (03): : 1261 - 1267
  • [2] CODON PREFERENCES IN FREE-LIVING MICROORGANISMS
    ANDERSSON, SGE
    KURLAND, CG
    [J]. MICROBIOLOGICAL REVIEWS, 1990, 54 (02) : 198 - 210
  • [3] [Anonymous], 1989, Chem Biochem Med Asp
  • [4] ELECTROBLOTTING OF INDIVIDUAL POLYPEPTIDES FROM SDS POLYACRYLAMIDE GELS FOR DIRECT SEQUENCE-ANALYSIS
    BERGMAN, T
    JORNVALL, H
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1987, 169 (01): : 9 - 12
  • [5] BJORNESTEDT R, 1992, BIOCHEM J, V282, P505
  • [6] IMPROVED SILVER STAINING OF PLANT-PROTEINS, RNA AND DNA IN POLYACRYLAMIDE GELS
    BLUM, H
    BEIER, H
    GROSS, HJ
    [J]. ELECTROPHORESIS, 1987, 8 (02) : 93 - 99
  • [7] BOARD PG, 1980, AM J HUM GENET, V32, P690
  • [8] BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
  • [9] S-(NITROCARBOBENZOXY)GLUTATHIONES - POTENT COMPETITIVE INHIBITORS OF MAMMALIAN GLYOXALASE-II
    BUSH, PE
    NORTON, SJ
    [J]. JOURNAL OF MEDICINAL CHEMISTRY, 1985, 28 (06) : 828 - 830
  • [10] INVIVO PROCESSING OF N-TERMINAL METHIONINE IN ESCHERICHIA-COLI
    DALBOGE, H
    BAYNE, S
    PEDERSEN, J
    [J]. FEBS LETTERS, 1990, 266 (1-2) : 1 - 3