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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4736732
- author
- 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. (More)
- Bielecka, Ewa
LU
; Scavenius, Carsten
; Kantyka, Tomasz
; Jusko, Monika
LU
; Mizgalska, Danuta
; Szmigielski, Borys
; Potempa, Barbara
; Enghild, Jan J
; Prossnitz, Eric
; Blom, Anna
LU
and Potempa, Jan
(Less)
- organization
- publishing date
- 2014
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 289
- issue
- 47
- pages
- 32481 - 32487
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- pmid:25324545
- wos:000345335000008
- scopus:84911906315
- pmid:25324545
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.C114.617142
- language
- English
- LU publication?
- yes
- id
- 341f1bd4-962f-485b-a608-de308f3ea61c (old id 4736732)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/25324545?dopt=Abstract
- date added to LUP
- 2016-04-01 11:02:19
- date last changed
- 2022-05-06 03:35:14
@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 = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{Peptidyl arginine deiminase from Porphyromonas gingivalis abolishes C5a activity.}},
url = {{https://lup.lub.lu.se/search/files/2330889/7584123.pdf}},
doi = {{10.1074/jbc.C114.617142}},
volume = {{289}},
year = {{2014}},
}