Expression of human leukotriene A4 hydrolase cDNA in Escherichia coli

FEBS Letters

Seyama, Yousuke¹  Kawasaki, Hiroshi²  Minami, Yasufumi²  Minami, Michiko¹  Ohno, Shigeo¹  Ohishi, Nobuya¹  Emori, Yasufumi²  Shimizu, Takao¹  Suzuki, Koichi² 

[1] Department of Physiological Chemistry and Nutrition, Faculty of Medicine, University of Tokyo, Tokyo 113, Japan;Department of Molecular Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 113, Japan

关键词: Leukotriene A4 hydrolase;    Leukotriene B4;    cDNA;    DNA expression;    (E. coli);    LT;    leukotriene;    LTA4;    (5S)-trans-5;    6-oxide-7;    9-trans-11;    14-cis-eicosatetraenoic acid;    LTB4;    (5S;    12R)-dihydroxy-6;    14-cis-8;    10-trans-eicosatetraenoic acid;    bp;    base pair;    IPTG;    isopropyl-1-thio-β-D-galactoside;    NEM;    N-ethylmaleimide;    PCMB;    p-chloromercuribenzoic acid;   

DOI  :  10.1016/0014-5793(88)81140-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The cDNA clone encoding human leukotriene A₄ hydrolase was inserted into a vector pUC9 and expressed in Escherichia coli as a fusion protein containing the first 10 amino acid residues derived from a vector. The leukotriene A₄ hydrolase activity was recovered in the soluble fraction of the transformants. The purified enzyme showed kinetic properties similar to the native enzyme, including inactivation by the substrate and sulfhydryl-modifying reagents. The results demonstrate that a protein with an M _r of 70000 was expressed in Escherichia coli with a full enzyme activity and structural fidelity. Acquisition of the expression system makes it feasible to elucidate the reaction mechanism of the enzyme.

【 授权许可】

Unknown