Expression and activity of citrullinating peptidylarginine deiminase enzymes in monocytes and macrophages.
BACKGROUND
Antibodies directed to proteins containing the non-standard amino acid citrulline, are extremely specific for rheumatoid arthritis (RA). Peptidylcitrulline can be generated by post-translational conversion of arginine residues. This process, citrullination, is catalysed by a group of calcium dependent peptidylarginine deiminase (PAD) enzymes.


OBJECTIVE
To investigate the expression and activity of four isotypes of PAD in peripheral blood and synovial fluid cells of patients with RA.


RESULTS
The data presented here show that citrullination of proteins by PAD enzymes is a process regulated at three levels: transcription-in peripheral blood PAD2 and PAD4 mRNAs are expressed predominantly in monocytes; PAD4 mRNA is not detectable in macrophages, translation-translation of PAD2 mRNA is subject to differentiation stage-specific regulation by its 3' UTR, and activation-the PAD proteins are only activated when sufficient Ca(2+) is available. Such high Ca(2+) concentrations are normally not present in living cells. In macrophages, which are abundant in the inflamed RA synovium, vimentin is specifically citrullinated after Ca(2+) influx.


CONCLUSIONS
PAD2 and PAD4 are the most likely candidate PAD isotypes for the citrullination of synovial proteins in RA. Our results indicate that citrullinated vimentin is a candidate autoantigen in RA.
Vossenaar
E R
ER
Department of Biochemistry, University of Nijmegen, Nijmegen, The Netherlands. e.vossenaar@ncmls.kun.nl
Radstake
T R D
TR
van der Heijden
A
A
van Mansum
M A M
MA
Dieteren
C
C
de Rooij
D-J
DJ
Barrera
P
P
Zendman
A J W
AJ
van Venrooij
W J
WJ
D001172:Arthritis, Rheumatoid / Q000201:enzymology* / Q000235:genetics; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D002956:Citrulline / Q000378:metabolism*; D006801:Humans; D006867:Hydrolases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008264:Macrophages / Q000201:enzymology*; D009000:Monocytes / Q000201:enzymology*; D014176:Protein Biosynthesis / Q000235:genetics; D000080064:Protein-Arginine Deiminase Type 2; D000076342:Protein-Arginine Deiminases; D012333:RNA, Messenger / Q000032:analysis; D013582:Synovial Fluid / Q000201:enzymology; D014158:Transcription, Genetic / Q000235:genetics; D014746:Vimentin / Q000276:immunology
D018950:Lipopolysaccharide Receptors; D012333:RNA, Messenger; D014746:Vimentin; D002956:Citrulline; D006867:Hydrolases; C000620352:PADI2 protein, human; D000080064:Protein-Arginine Deiminase Type 2; D000076342:Protein-Arginine Deiminases; D002118:Calcium