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Summary of citrullinated proteins as candidate autoantigens in RA

Summary of citrullinated proteins as candidate autoantigens in RA. Fibrin(ogen). human. BBRC 327(1):192-200 (2005)*. J Immunol.166(6):4177-84(2001). Filaggrin. human. BBRC 327(1):192-200 (2005)*. J Immunol.162(1):585-94 (1999). eukaryotic translation initiation factor 4G1 (eIF4G1).

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Summary of citrullinated proteins as candidate autoantigens in RA

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  1. Summary of citrullinated proteins as candidate autoantigens in RA Fibrin(ogen) human BBRC 327(1):192-200 (2005)* J Immunol.166(6):4177-84(2001) Filaggrin human BBRC 327(1):192-200 (2005)* J Immunol.162(1):585-94 (1999) eukaryotic translation initiation factor 4G1 (eIF4G1) human In press* Antithrombin III human Rheumatology 44(3):293-298(2005)* Fibronectin human Rheumatology 44(11):1374-1382(2005)* human BBRC 333(2):418-26 (2005)* Type I collagen Type II collagen human, rat Eur J Immunol. 35(5):1643-52 (2005) Vimentin human Arthritis Res Ther. 6(2):R142-50(2004) a-Enolase human Arthritis Res Ther. 7(6):R1421-9(2005) * Reported by RIKEN

  2. Citrullination of fibrinogen by PADI4 1. CBB staining 2. Western blotting + + + + + + Fibrinogen PADI4 33mM EDTA Fibrinogen PADI4 33mM EDTA + - + - + + - - + - - + CBB staining 83kDa Citrullinated Fibrinogen A a 66 kDa B b 56 kDa 41kDa g 51 kDa a-modified citrulline hPADI4 Concentration of citrulline (uM) PADI4 Vmax (mmol/mg/hr) =33.2 Km (mM) =0.055 No enzyme min

  3. 0.06 0.05 0.04 0.03 0.02 0.01 0 -0.01 min 0 10 20 30 40 50 The inhibition of fibrin polymerization by PADI4 OD350nm OD350nm 0.04 0.03 0.02 0.01 0 -0.01 min 0 10 20 30 40 50 : fibrinogen only :citrullinated fibrinogen+EDTA : fibrinogen : citrullinated fibrinogen = 9 : 1 :native fibrinogen+EDTA :citrullinated fibrinogen+Ca : fibrinogen : citrullinated fibrinogen = 8 : 2 :native fibrinogen+Ca : fibrinogen : citrullinated fibrinogen = 7 : 3 : fibrinogen : citrullinated fibrinogen = 5 : 5

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

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