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Peptidyl arginine deiminase from Porphyromonas gingivalis abolishes C5a activity.

Bielecka, Ewa LU ; Scavenius, Carsten; Kantyka, Tomasz; Jusko, Monika LU ; Mizgalska, Danuta; Szmigielski, Borys; Potempa, Barbara; Enghild, Jan J; Prossnitz, Eric and Blom, Anna LU , et al. (2014) In Journal of Biological Chemistry 289(47). p.32481-32487
Abstract
Evasion of killing by the complement system, a crucial part of innate immunity, is a key evolutionary strategy of many human pathogens. A major etiological agent of chronic periodontitis, the Gram-negative bacterium Porphyromonas gingivalis produces a vast arsenal of virulence factors that compromise human defense mechanisms. One of these is peptidylarginine deiminase (PPAD), an enzyme unique to P. gingivalis among bacteria, which converts Arg residues in polypeptide chains into citrulline. Here, we report that PPAD citrullination of a critical C-terminal arginine of the anaphylatoxin C5a disabled the protein function. Treatment of C5a with PPAD in vitro resulted in decreased chemotaxis of human neutrophils and diminished calcium signaling... (More)
Evasion of killing by the complement system, a crucial part of innate immunity, is a key evolutionary strategy of many human pathogens. A major etiological agent of chronic periodontitis, the Gram-negative bacterium Porphyromonas gingivalis produces a vast arsenal of virulence factors that compromise human defense mechanisms. One of these is peptidylarginine deiminase (PPAD), an enzyme unique to P. gingivalis among bacteria, which converts Arg residues in polypeptide chains into citrulline. Here, we report that PPAD citrullination of a critical C-terminal arginine of the anaphylatoxin C5a disabled the protein function. Treatment of C5a with PPAD in vitro resulted in decreased chemotaxis of human neutrophils and diminished calcium signaling in monocytic cell line U937 transfected with the C5a receptor (C5aR) and loaded with a fluorescent intracellular calcium probe: Fura 2-AM. Moreover, a low degree of citrullination of internal arginine residues by PPAD was also detected using mass spectrometry. Further, after treatment of C5 with outer membrane vesicles (OMVs) naturally shed by P. gingivalis we observed generation of C5a totally citrullinated at the C-terminal Arg74 residue (Arg74Cit). In stark contrast only native C5a was detected after treatment with PPAD-null OMVs. Our study suggests reduced antibacterial and proinflammatory capacity of citrullinated C5a, achieved via lower level of chemotactic potential of the modified molecule, and weaker cell activation. In the context of previous studies, which showed crosstalk between C5aR and toll-like receptors, as well as enhanced arthritis development in mice infected with PPAD expressing P. gingivalis, our findings support a crucial role of PPAD in the virulence of P. gingivalis. (Less)
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Journal of Biological Chemistry
volume
289
issue
47
pages
32481 - 32487
publisher
ASBMB
external identifiers
  • pmid:25324545
  • wos:000345335000008
  • scopus:84911906315
ISSN
1083-351X
DOI
10.1074/jbc.C114.617142
language
English
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yes
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341f1bd4-962f-485b-a608-de308f3ea61c (old id 4736732)
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http://www.ncbi.nlm.nih.gov/pubmed/25324545?dopt=Abstract
date added to LUP
2014-11-09 21:53:05
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2017-11-05 03:23:12
@article{341f1bd4-962f-485b-a608-de308f3ea61c,
  abstract     = {Evasion of killing by the complement system, a crucial part of innate immunity, is a key evolutionary strategy of many human pathogens. A major etiological agent of chronic periodontitis, the Gram-negative bacterium Porphyromonas gingivalis produces a vast arsenal of virulence factors that compromise human defense mechanisms. One of these is peptidylarginine deiminase (PPAD), an enzyme unique to P. gingivalis among bacteria, which converts Arg residues in polypeptide chains into citrulline. Here, we report that PPAD citrullination of a critical C-terminal arginine of the anaphylatoxin C5a disabled the protein function. Treatment of C5a with PPAD in vitro resulted in decreased chemotaxis of human neutrophils and diminished calcium signaling in monocytic cell line U937 transfected with the C5a receptor (C5aR) and loaded with a fluorescent intracellular calcium probe: Fura 2-AM. Moreover, a low degree of citrullination of internal arginine residues by PPAD was also detected using mass spectrometry. Further, after treatment of C5 with outer membrane vesicles (OMVs) naturally shed by P. gingivalis we observed generation of C5a totally citrullinated at the C-terminal Arg74 residue (Arg74Cit). In stark contrast only native C5a was detected after treatment with PPAD-null OMVs. Our study suggests reduced antibacterial and proinflammatory capacity of citrullinated C5a, achieved via lower level of chemotactic potential of the modified molecule, and weaker cell activation. In the context of previous studies, which showed crosstalk between C5aR and toll-like receptors, as well as enhanced arthritis development in mice infected with PPAD expressing P. gingivalis, our findings support a crucial role of PPAD in the virulence of P. gingivalis.},
  author       = {Bielecka, Ewa and Scavenius, Carsten and Kantyka, Tomasz and Jusko, Monika and Mizgalska, Danuta and Szmigielski, Borys and Potempa, Barbara and Enghild, Jan J and Prossnitz, Eric and Blom, Anna and Potempa, Jan},
  issn         = {1083-351X},
  language     = {eng},
  number       = {47},
  pages        = {32481--32487},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Peptidyl arginine deiminase from Porphyromonas gingivalis abolishes C5a activity.},
  url          = {http://dx.doi.org/10.1074/jbc.C114.617142},
  volume       = {289},
  year         = {2014},
}