Thiamine biosynthesis in Escherichia coli: isolation and initial characterisation of the ThiGH complex

FEBS Letters

Lowe, David J²  Roach, Peter L¹  Kriek, Marco¹  Leonardi, Roberta¹  Fairhurst, Shirley A² 

[1]Department of Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ, UK

[2]Biological Chemistry Department, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK

关键词: Thiamine biosynthesis;    Iron–sulfur protein;    Electron paramagnetic resonance;    ThiH-His;    ThiH with a (His)6 tag attached to the C-terminus;    EPR;    electron paramagnetic resonance;    BSA;    bovine serum albumin;    DTT;    dithiothreitol;    PLP;    pyridoxal phosphate;   

DOI  :  10.1016/S0014-5793(03)00204-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In Escherichia coli, two of the proteins required for the biosynthesis of the thiazole moiety of thiamine (vitamin B₁) are ThiG and ThiH, encoded as part of the thiCEFSGH operon. In this study, a C-terminally hexahistidine-tagged ThiH (ThiH-His) was expressed in E. coli as a soluble protein from thiGH-His-tag and thiFSGH-His-tag-bearing plasmids. When isolated under anaerobic conditions, ThiG and ThiH-His co-purify as a large multimeric non-covalent complex. Electron paramagnetic resonance and UV–visible spectroscopy together with iron and sulfide analyses revealed the presence of an iron–sulfur cluster within this complex.

【 授权许可】

Unknown