Generation of SUMO‐1 modified proteins in E. coli: towards understanding the biochemistry/structural biology of the SUMO‐1 pathway

FEBS Letters

Uchimura, Yasuhiro¹  Saitoh, Hisato¹  Nakao, Mitsuyoshi¹ 

[1] Department of Regeneration Medicine, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan

关键词: Posttranslational modification;    SUMO-1;    Ran GTPase activating protein 1;    Ran binding protein 2;    Promyelocytic leukemia;    p53;    SUMO-1;    small ubiquitin-related modifier-1;    Ubc9;    ubiquitin conjugating enzyme 9;    RanGAP1;    Ran GTPase activating protein 1;    RanBP2;    Ran binding protein 2;    PML;    promyelocytic leukemia;    SDS;    sodium dodecyl sulfate;    IPTG;    isopropyl-β-D-thiogalactopyranoside;    DTT;    dithiothreitol;   

DOI  :  10.1016/S0014-5793(04)00321-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Here, we developed a binary vector system that introduces a synthetic SUMO-1 conjugation pathway into Escherichia coli and demonstrated that large amounts of sumoylated Ran GTPase activating protein 1 C-terminal region (RanGAP1-C2), Ran binding protein 2 internal repeat domain, p53 and promyelocytic leukemia were efficiently produced. The sumoylated recombinant RanGAP1-C2 appeared to retain the in vivo properties, since it was specifically sumoylated at lysine 517 as expected from in vivo studies. Our findings indicate the establishment of a biosynthetic route for producing large amounts of sumoylated recombinant proteins that will open up new avenues for studying the biochemical and structural aspects of the SUMO-1 modification pathway.

【 授权许可】

Unknown