Summary of citrullinated proteins as candidate autoantigens in

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 4 G 1 (e. IF 4 G 1) human In press* Antithrombin III human Rheumatology 44(3): 293 -298(2005)* Fibronectin human Rheumatology 44(11): 1374 -1382(2005)* Type I collagen human BBRC 333(2): 418 -26 (2005)* Type II collagen human, rat Eur J Immunol. 35(5): 1643 -52 (2005) Vimentin human Arthritis Res Ther. 6(2): R 142 -50(2004) a-Enolase human Arthritis Res Ther. 7(6): R 1421 -9(2005) * Reported by RIKEN

Citrullination of fibrinogen by PADI 4 1. CBB staining Fibrinogen PADI 4 33 m. M EDTA 2. Western blotting + + - + + + + + - CBB+staining Fibrinogen PADI 4 33 m. M EDTA 83 k. Da A a 66 k. Da B b 56 k. Da g 51 k. Da Citrullinated Fibrinogen 41 k. Da Concentration of citrulline (u. M) a-modified citrulline h. PADI 4 No enzyme min PADI 4 Vmax (mmol/mg/hr) =33. 2 Km (m. M) =0. 055

The inhibition of fibrin polymerization by PADI 4 OD 350 nm 0. 06 0. 04 0. 05 0. 03 0. 04 0. 02 0. 03 0. 02 0. 01 0 0 -0. 01 0 10 20 30 40 : citrullinated fibrinogen+EDTA : native fibrinogen+EDTA : citrullinated fibrinogen+Ca : native fibrinogen+Ca 50 min -0. 01 0 10 20 30 40 50 min : fibrinogen only : fibrinogen : citrullinated fibrinogen = 9 : 1 : fibrinogen : citrullinated fibrinogen = 8 : 2 : fibrinogen : citrullinated fibrinogen = 7 : 3 : fibrinogen : citrullinated fibrinogen = 5 : 5

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