Thiamine biosynthesis in Escherichia coli: isolation and initial characterisation of the ThiGH complex
Thiamine biosynthesis in Escherichia coli: isolation and initial characterisation of the ThiGH complex
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.
thiamine biosynthesis, iron-sulfur protein, electron paramagnetic, resonancein-vitro biosynthesis, biotin synthase, sulfane sulfur, transfer-rna, iron, protein, identification, 4-thiouridine, cluster, enzyme
95-99
Leonardi, Roberta
d5419843-b210-4d01-a76e-c5a8fceed358
Fairhurst, Shirley A.
fd1d174b-14ae-461c-b15c-d406f1453b77
Kriek, Marco
c3e820c2-c59b-49c8-a792-f64a69e4401a
Lowe, David J.
aa0e800a-20e9-4c09-b8d3-3256a8079e3f
Roach, Peter L.
ca94060c-4443-482b-af3e-979243488ba9
27 March 2003
Leonardi, Roberta
d5419843-b210-4d01-a76e-c5a8fceed358
Fairhurst, Shirley A.
fd1d174b-14ae-461c-b15c-d406f1453b77
Kriek, Marco
c3e820c2-c59b-49c8-a792-f64a69e4401a
Lowe, David J.
aa0e800a-20e9-4c09-b8d3-3256a8079e3f
Roach, Peter L.
ca94060c-4443-482b-af3e-979243488ba9
Leonardi, Roberta, Fairhurst, Shirley A., Kriek, Marco, Lowe, David J. and Roach, Peter L.
(2003)
Thiamine biosynthesis in Escherichia coli: isolation and initial characterisation of the ThiGH complex.
FEBS Letters, 539 (1-3), .
(doi: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.
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Published date: 27 March 2003
Keywords:
thiamine biosynthesis, iron-sulfur protein, electron paramagnetic, resonancein-vitro biosynthesis, biotin synthase, sulfane sulfur, transfer-rna, iron, protein, identification, 4-thiouridine, cluster, enzyme
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Local EPrints ID: 20018
URI: http://eprints.soton.ac.uk/id/eprint/20018
ISSN: 0014-5793
PURE UUID: 039a5ae9-b59c-45bf-a52e-ed3cc111af28
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Date deposited: 24 Feb 2006
Last modified: 15 Mar 2024 06:21
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Contributors
Author:
Roberta Leonardi
Author:
Shirley A. Fairhurst
Author:
Marco Kriek
Author:
David J. Lowe
Author:
Peter L. Roach
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