The peroxidase and peroxynitrite reductase activity of human erythrocyte peroxiredoxin 2.
Peroxiredoxin 2 (Prx2) is a 2-Cys peroxiredoxin extremely abundant in the erythrocyte. The peroxidase activity was studied in a steady-state approach yielding an apparent K(M) of 2.4 microM for human thioredoxin and a very low K(M) for H2O2 (0.7 microM). Rate constants for the reaction of peroxidatic cysteine with the peroxide substrate, H2O2 or peroxynitrite, were determined by competition kinetics, k(2) = 1.0 x 10(8) and 1.4 x 10(7) M(-1) s(-1) at 25 degrees C and pH 7.4, respectively. Excess of both oxidants inactivated the enzyme by overoxidation and also tyrosine nitration and dityrosine were observed with peroxynitrite treatment. Prx2 associates into decamers (5 homodimers) and we estimated a dissociation constant K(d) < 10(-23) M(4) which confirms the enzyme exists as a decamer in vivo. Our kinetic results indicate Prx2 is a key antioxidant enzyme for the erythrocyte and reveal red blood cells as active oxidant scrubbers in the bloodstream.
Manta
Bruno
B
Laboratorio de Fisicoquímica Biológica, Instituto de Química Biológica, Facultad de Ciencias, Universidad de la República, Iguá 4225, 11400 Montevideo, Uruguay.
Hugo
Martín
M
Ortiz
Cecilia
C
Ferrer-Sueta
Gerardo
G
Trujillo
Madia
M
Denicola
Ana
A
D000818:Animals; D002384:Catalysis; D002850:Chromatography, Gel; D019281:Dimerization; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000097:blood / Q000378:metabolism; D007700:Kinetics; D008322:Mammals; D009249:NADP / Q000097:blood; D016877:Oxidants / Q000097:blood; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000097:blood; D010544:Peroxidases / Q000097:blood; D054464:Peroxiredoxins / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D013879:Thioredoxins / Q000097:blood
D016877:Oxidants; D013879:Thioredoxins; D009249:NADP; D006861:Hydrogen Peroxide; D010088:Oxidoreductases; D010544:Peroxidases; C086065:PRDX2 protein, human; D054464:Peroxiredoxins; C109110:peroxynitrite reductase