Summary of citrullinated proteins as candidate autoantigens in RA Fibrin(ogen) Filaggrin Type I collagen eukaryotic translation initiation factor 4G1 (eIF4G1)

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Summary of citrullinated proteins as candidate autoantigens in RA Fibrin(ogen) Filaggrin Type I collagen eukaryotic translation initiation factor 4G1 (eIF4G1) Antithrombin III In press* Rheumatology 44(3): (2005)* FibronectinRheumatology 44(11): (2005)* Type II collagen human human, rat BBRC 327(1): (2005)* J Immunol.166(6): (2001) J Immunol.162(1): (1999) BBRC 333(2): (2005)* * Reported by RIKEN VimentinhumanArthritis Res Ther. 6(2):R142-50(2004)  -Enolase human Arthritis Res Ther. 7(6):R1421-9(2005) Eur J Immunol. 35(5): (2005)

Citrullination of fibrinogen by PADI4 min Concentration of citrulline (uM) hPADI4 No enzyme PADI4 Vmax (mmol/mg/hr) =33.2 Km (mM) =0.055 B  56 kDa  51 kDa A  66 kDa CBB staining Fibrinogen PADI4 33mM EDTA kDa 41kDa Citrullinated Fibrinogen 2. Western blotting  -modified citrulline Fibrinogen PADI4 33mM EDTA CBB staining

: fibrinogen only : fibrinogen : citrullinated fibrinogen = 9 : 1 : fibrinogen : citrullinated fibrinogen = 8 : 2 : fibrinogen : citrullinated fibrinogen = 7 : 3 : fibrinogen : citrullinated fibrinogen = 5 : min OD350nm The inhibition of fibrin polymerization by PADI4 :citrullinated fibrinogen+EDTA :native fibrinogen+EDTA :citrullinated fibrinogen+Ca :native fibrinogen+Ca OD350nm

Positive control Human FGA PADI4+ PADI4- Synoviocyte cDNA library (2 nd ) Citrullinated Uncitrullinated - + PADI Citrullinated - + PADI Type I collagen C IC II C IC II CBB  -MC (1:1000) Identification and confirmation of citrullinated CI modified by PADIs