The active-site cysteinyls and hydrophobic cavity residues of resA are important for cytochrome c maturation in Bacillus subtilis
(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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https://lup.lub.lu.se/record/1209087
- author
- Hodson, Christopher ; Lewin, Allison ; Hederstedt, Lars LU and Le Brun, Nick
- organization
- publishing date
- 2008
- 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
- 2016-04-01 11:51:21
- date last changed
- 2022-01-26 19:14:49
@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}},
doi = {{10.1128/JB.00145-08}},
volume = {{190}},
year = {{2008}},
}