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Structure-function analysis of PrsA reveals roles for the parvulin-like and flanking N- and C-terminal domains in protein folding and secretion in Bacillus subtilis.

Vitikainen, Marika; Lappalainen, Ilkka; Seppala, Raili; Antelmann, Haike; Boer, Harry; Taira, Suvi; Savilahti, Harri; Hecker, Michael; Vihinen, Mauno LU and Sarvas, Matti, et al. (2004) In Journal of Biological Chemistry 279(18). p.19302-19314
Abstract
The PrsA protein of Bacillus subtilis is an essential membrane-bound lipoprotein that is assumed to assist post-translocational folding of exported proteins and stabilize them in the compartment between the cytoplasmic membrane and cell wall. This folding activity is consistent with the homology of a segment of PrsA with parvulin-type peptidyl-prolyl cis/trans isomerases (PPIase). In this study, molecular modeling showed that the parvulin-like region can adopt a parvulin-type fold with structurally conserved active site residues. PrsA exhibits PPIase activity in a manner dependent on the parvulin-like domain. We constructed deletion, peptide insertion, and amino acid substitution mutations and demonstrated that the parvulin-like domain as... (More)
The PrsA protein of Bacillus subtilis is an essential membrane-bound lipoprotein that is assumed to assist post-translocational folding of exported proteins and stabilize them in the compartment between the cytoplasmic membrane and cell wall. This folding activity is consistent with the homology of a segment of PrsA with parvulin-type peptidyl-prolyl cis/trans isomerases (PPIase). In this study, molecular modeling showed that the parvulin-like region can adopt a parvulin-type fold with structurally conserved active site residues. PrsA exhibits PPIase activity in a manner dependent on the parvulin-like domain. We constructed deletion, peptide insertion, and amino acid substitution mutations and demonstrated that the parvulin-like domain as well as flanking N- and C-terminal domains are essential for in vivo PrsA function in protein secretion and growth. Surprisingly, none of the predicted active site residues of the parvulin-like domain was essential for growth and protein secretion, although several active site mutations reduced or abolished the PPIase activity or the ability of PrsA to catalyze proline-limited protein folding in vitro. Our results indicate that PrsA is a PPIase, but the essential role in vivo seems to depend on some non-PPIase activity of both the parvulin-like and flanking domains. (Less)
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keywords
Bacillus subtilis: chemistry, Bacillus subtilis: metabolism, Bacterial Proteins: chemistry, Lipoproteins: chemistry, Lipoproteins: genetics, Lipoproteins: physiology, Membrane Proteins: chemistry, Membrane Proteins: genetics, Membrane Proteins: physiology, Peptidylprolyl Isomerase: chemistry, Proteins: secretion
in
Journal of Biological Chemistry
volume
279
issue
18
pages
19302 - 19314
publisher
ASBMB
external identifiers
  • pmid:14976191
  • scopus:2442446271
ISSN
1083-351X
DOI
10.1074/jbc.M400861200
language
English
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no
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df11a346-c679-456b-906f-9b491c0ce355 (old id 3635496)
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http://www.ncbi.nlm.nih.gov/pubmed/14976191?dopt=Abstract
date added to LUP
2013-06-12 16:07:59
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2017-08-20 04:36:45
@article{df11a346-c679-456b-906f-9b491c0ce355,
  abstract     = {The PrsA protein of Bacillus subtilis is an essential membrane-bound lipoprotein that is assumed to assist post-translocational folding of exported proteins and stabilize them in the compartment between the cytoplasmic membrane and cell wall. This folding activity is consistent with the homology of a segment of PrsA with parvulin-type peptidyl-prolyl cis/trans isomerases (PPIase). In this study, molecular modeling showed that the parvulin-like region can adopt a parvulin-type fold with structurally conserved active site residues. PrsA exhibits PPIase activity in a manner dependent on the parvulin-like domain. We constructed deletion, peptide insertion, and amino acid substitution mutations and demonstrated that the parvulin-like domain as well as flanking N- and C-terminal domains are essential for in vivo PrsA function in protein secretion and growth. Surprisingly, none of the predicted active site residues of the parvulin-like domain was essential for growth and protein secretion, although several active site mutations reduced or abolished the PPIase activity or the ability of PrsA to catalyze proline-limited protein folding in vitro. Our results indicate that PrsA is a PPIase, but the essential role in vivo seems to depend on some non-PPIase activity of both the parvulin-like and flanking domains.},
  author       = {Vitikainen, Marika and Lappalainen, Ilkka and Seppala, Raili and Antelmann, Haike and Boer, Harry and Taira, Suvi and Savilahti, Harri and Hecker, Michael and Vihinen, Mauno and Sarvas, Matti and Kontinen, Vesa P},
  issn         = {1083-351X},
  keyword      = {Bacillus subtilis: chemistry,Bacillus subtilis: metabolism,Bacterial Proteins: chemistry,Lipoproteins: chemistry,Lipoproteins: genetics,Lipoproteins: physiology,Membrane Proteins: chemistry,Membrane Proteins: genetics,Membrane Proteins: physiology,Peptidylprolyl Isomerase: chemistry,Proteins: secretion},
  language     = {eng},
  number       = {18},
  pages        = {19302--19314},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Structure-function analysis of PrsA reveals roles for the parvulin-like and flanking N- and C-terminal domains in protein folding and secretion in Bacillus subtilis.},
  url          = {http://dx.doi.org/10.1074/jbc.M400861200},
  volume       = {279},
  year         = {2004},
}