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Production, purification and detergent exchange of isotopically labeled Bacillus subtilis cytochrome b(558) (SdhC)

Baureder, Michael LU and Hederstedt, Lars LU (2011) In Protein Expression and Purification 80(1). p.97-101
Abstract
Cytochrome 6558 of the gram-positive bacterium Bacillus subtilis is the membrane anchor subunit of the succinate:quinone oxidoreductase of the citric acid cycle. The cytochrome consists of the SdhC polypeptide (202 residues) and two protoheme IX groups that function in transmembrane electron transfer to menaquinone. The general structure of the cytochrome is known from extensive experimental studies and by comparison to Wolinella succinogenes fumarate reductase for which the X-ray crystal structure has been determined. Solution state NMR can potentially be used to identify the quinone binding site(s) and study, e.g. redox-linked, dynamics of cytochrome b(558). In this work we present an efficient procedure for the isolation of preparative... (More)
Cytochrome 6558 of the gram-positive bacterium Bacillus subtilis is the membrane anchor subunit of the succinate:quinone oxidoreductase of the citric acid cycle. The cytochrome consists of the SdhC polypeptide (202 residues) and two protoheme IX groups that function in transmembrane electron transfer to menaquinone. The general structure of the cytochrome is known from extensive experimental studies and by comparison to Wolinella succinogenes fumarate reductase for which the X-ray crystal structure has been determined. Solution state NMR can potentially be used to identify the quinone binding site(s) and study, e.g. redox-linked, dynamics of cytochrome b(558). In this work we present an efficient procedure for the isolation of preparative amounts of isotopically labeled B. subtilis cytochrome 6558 produced in Escherichia coli. We have also evaluated several detergents suitable for NMR for their effectiveness in maintaining the cytochrome solubilized and intact for days at room temperature. (C) 2011 Elsevier Inc. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Bacillus subtilis, Succinate dehydrogenase, Cytochrome, NMR, Detergent, exchange, Isotopic labeling
in
Protein Expression and Purification
volume
80
issue
1
pages
97 - 101
publisher
Academic Press
external identifiers
  • wos:000295608800015
  • pmid:21641999
  • scopus:80053310871
ISSN
1046-5928
DOI
10.1016/j.pep.2011.05.013
language
English
LU publication?
yes
id
ac42e6d4-6192-4872-9fb3-a62114e37e05 (old id 2212208)
date added to LUP
2011-11-24 13:22:31
date last changed
2017-08-09 14:41:31
@article{ac42e6d4-6192-4872-9fb3-a62114e37e05,
  abstract     = {Cytochrome 6558 of the gram-positive bacterium Bacillus subtilis is the membrane anchor subunit of the succinate:quinone oxidoreductase of the citric acid cycle. The cytochrome consists of the SdhC polypeptide (202 residues) and two protoheme IX groups that function in transmembrane electron transfer to menaquinone. The general structure of the cytochrome is known from extensive experimental studies and by comparison to Wolinella succinogenes fumarate reductase for which the X-ray crystal structure has been determined. Solution state NMR can potentially be used to identify the quinone binding site(s) and study, e.g. redox-linked, dynamics of cytochrome b(558). In this work we present an efficient procedure for the isolation of preparative amounts of isotopically labeled B. subtilis cytochrome 6558 produced in Escherichia coli. We have also evaluated several detergents suitable for NMR for their effectiveness in maintaining the cytochrome solubilized and intact for days at room temperature. (C) 2011 Elsevier Inc. All rights reserved.},
  author       = {Baureder, Michael and Hederstedt, Lars},
  issn         = {1046-5928},
  keyword      = {Bacillus subtilis,Succinate dehydrogenase,Cytochrome,NMR,Detergent,exchange,Isotopic labeling},
  language     = {eng},
  number       = {1},
  pages        = {97--101},
  publisher    = {Academic Press},
  series       = {Protein Expression and Purification},
  title        = {Production, purification and detergent exchange of isotopically labeled <em>Bacillus subtilis </em>cytochrome b(558) (SdhC)},
  url          = {http://dx.doi.org/10.1016/j.pep.2011.05.013},
  volume       = {80},
  year         = {2011},
}