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

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Author

Summary, in English

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.

Publishing year

2011

Language

English

Pages

97-101

Publication/Series

Protein Expression and Purification

Volume

80

Issue

1

Document type

Journal article

Publisher

Academic Press

Topic

  • Biological Sciences
  • Microbiology

Keywords

  • Bacillus subtilis
  • Succinate dehydrogenase
  • Cytochrome
  • NMR
  • Detergent
  • exchange
  • Isotopic labeling

Status

Published

ISBN/ISSN/Other

  • ISSN: 1046-5928