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The active-site cysteinyls and hydrophobic cavity residues of resA are important for cytochrome c maturation in Bacillus subtilis

Hodson, Christopher; Lewin, Allison; Hederstedt, Lars LU and Le Brun, Nick (2008) In Journal of Bacteriology 190(13). p.4697-4705
Abstract
ResA is an extracytoplasmic membrane-bound thiol-disulfide oxidoreductase required for cytochrome c maturation in Bacillus subtilis. Previous biochemical and structural studies have revealed that the active-site cysteinyls cycle between oxidized and reduced states with a low reduction potential and that, upon reduction, a hydrophobic cavity forms close to the active site. Here we report in vivo studies of ResA-deficient B. subtilis complemented with a series of ResA variants. Using a range of methods to analyze the cellular cytochrome c content, we demonstrated (i) that the N-terminal transmembrane segment of ResA serves principally to anchor the protein to the cytoplasmic membrane but also plays a role in mediating the activity of the... (More)
ResA is an extracytoplasmic membrane-bound thiol-disulfide oxidoreductase required for cytochrome c maturation in Bacillus subtilis. Previous biochemical and structural studies have revealed that the active-site cysteinyls cycle between oxidized and reduced states with a low reduction potential and that, upon reduction, a hydrophobic cavity forms close to the active site. Here we report in vivo studies of ResA-deficient B. subtilis complemented with a series of ResA variants. Using a range of methods to analyze the cellular cytochrome c content, we demonstrated (i) that the N-terminal transmembrane segment of ResA serves principally to anchor the protein to the cytoplasmic membrane but also plays a role in mediating the activity of the protein; (ii) that the active-site cysteines are important for cytochrome c maturation activity; (iii) that Pro141, which forms part of the hydrophobic cavity and which adopts a cis conformation, plays an important role in protein stability; (iv) that Glu80, which lies at the base of the hydrophobic cavity, is important for cytochrome c maturation activity; and, finally, (v) that Pro141 and Glu80 ResA mutant variants promote selective maturation of low levels of one c-type cytochrome, subunit II of the cytochrome c oxidase caa(3), indicating that this apocytochrome is distinct from the other three endogenous c-type cytochromes of B. subtilis. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Bacteriology
volume
190
issue
13
pages
4697 - 4705
publisher
American Society for Microbiology
external identifiers
  • wos:000257174400036
  • scopus:46049093973
ISSN
0021-9193
DOI
10.1128/JB.00145-08
language
English
LU publication?
yes
id
4eb1e6f0-9d73-4f00-8381-ebfef01cd1c4 (old id 1209087)
date added to LUP
2008-09-04 09:38:11
date last changed
2017-08-09 16:08:43
@article{4eb1e6f0-9d73-4f00-8381-ebfef01cd1c4,
  abstract     = {ResA is an extracytoplasmic membrane-bound thiol-disulfide oxidoreductase required for cytochrome c maturation in Bacillus subtilis. Previous biochemical and structural studies have revealed that the active-site cysteinyls cycle between oxidized and reduced states with a low reduction potential and that, upon reduction, a hydrophobic cavity forms close to the active site. Here we report in vivo studies of ResA-deficient B. subtilis complemented with a series of ResA variants. Using a range of methods to analyze the cellular cytochrome c content, we demonstrated (i) that the N-terminal transmembrane segment of ResA serves principally to anchor the protein to the cytoplasmic membrane but also plays a role in mediating the activity of the protein; (ii) that the active-site cysteines are important for cytochrome c maturation activity; (iii) that Pro141, which forms part of the hydrophobic cavity and which adopts a cis conformation, plays an important role in protein stability; (iv) that Glu80, which lies at the base of the hydrophobic cavity, is important for cytochrome c maturation activity; and, finally, (v) that Pro141 and Glu80 ResA mutant variants promote selective maturation of low levels of one c-type cytochrome, subunit II of the cytochrome c oxidase caa(3), indicating that this apocytochrome is distinct from the other three endogenous c-type cytochromes of B. subtilis.},
  author       = {Hodson, Christopher and Lewin, Allison and Hederstedt, Lars and Le Brun, Nick},
  issn         = {0021-9193},
  language     = {eng},
  number       = {13},
  pages        = {4697--4705},
  publisher    = {American Society for Microbiology},
  series       = {Journal of Bacteriology},
  title        = {The active-site cysteinyls and hydrophobic cavity residues of resA are important for cytochrome c maturation in <em>Bacillus subtilis</em>},
  url          = {http://dx.doi.org/10.1128/JB.00145-08},
  volume       = {190},
  year         = {2008},
}