Thiamine biosynthesis in Escherichia coli: isolation and initial characterisation of the ThiGH complex
- 6 March 2003
- journal article
- Published by Wiley in FEBS Letters
- Vol. 539 (1-3) , 95-99
- https://doi.org/10.1016/s0014-5793(03)00204-7
Abstract
In Escherichia coli, two of the proteins required for the biosynthesis of the thiazole moiety of thiamine (vitamin B(1)) 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.Keywords
This publication has 29 references indexed in Scilit:
- Biotin Synthase Is a Pyridoxal Phosphate-Dependent Cysteine DesulfuraseBiochemistry, 2002
- Enzymatic Modification of tRNAsJournal of Biological Chemistry, 2002
- Lesions in gshA (Encoding γ- l -Glutamyl- l -Cysteine Synthetase) Prevent Aerobic Synthesis of Thiamine in Salmonella enterica Serovar Typhimurium LT2Journal of Bacteriology, 2000
- Pyruvate Formate-Lyase-Activating Enzyme: Strictly Anaerobic Isolation Yields Active Enzyme Containing a [3Fe–4S]+ ClusterBiochemical and Biophysical Research Communications, 2000
- Evidence That ThiI, an Enzyme Shared between Thiamin and 4-Thiouridine Biosynthesis, May Be a Sulfurtransferase That Proceeds through a Persulfide IntermediateJournal of Biological Chemistry, 2000
- IscS Is a Sulfurtransferase for the in Vitro Biosynthesis of 4-Thiouridine in Escherichia coli tRNABiochemistry, 1999
- Efficient sequence analysis of the six gene products (7‐74 kDA) from the escherichia coli thiamin biosynthetic operon by tandem high‐resolution mass spectrometryProtein Science, 1998
- Thiamin Biosynthesis in Escherichia coli: IDENTIFICATION OF ThiS THIOCARBOXYLATE AS THE IMMEDIATE SULFUR DONOR IN THE THIAZOLE FORMATIONPublished by Elsevier ,1998
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976
- Über die Synthese des antineuritischen VitaminsBerichte der deutschen chemischen Gesellschaft (A and B Series), 1937