Title of article :
Reactions of yeast thioredoxin peroxidases I and II with hydrogen peroxide and peroxynitrite: Rate constants by competitive kinetics
Author/Authors :
Renata Ogusucu، نويسنده , , Daniel Rettori، نويسنده , , Daniela Cristina Munhoz، نويسنده , , Luis Eduardo Soares Netto، نويسنده , , Ohara Augusto، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2007
Abstract :
Peroxiredoxins are receiving increasing attention as defenders against oxidative damage and sensors of hydrogen peroxide-mediated signaling events. Likely to be critical for both functions is a rapid reaction with hydrogen peroxide, typically with second-order rate constants higher than 105 M−1 s−1. Until recently, however, the values reported for these rate constants have been in the range of 104−105 M−1 s−1, including those for cytosolic thioredoxin peroxidases I (Tsa1) and II (Tsa2) from Saccharomyces cerevisiae. To resolve this apparent paradox, we developed a competitive kinetic approach with horseradish peroxidase to determine the second-order rate constant of the reaction of peroxiredoxins with peroxynitrite and hydrogen peroxide. This method was validated and allowed for the determination of the second-order rate constant of the reaction of Tsa1 and Tsa2 with peroxynitrite (k 105 M−1 s−1) and hydrogen peroxide (k 107 M−1 s−1) at pH 7.4, 25°C. It also permitted the determination of the pKa of the peroxidatic cysteine of Tsa1 and Tsa2 (Cys47) as 5.4 and 6.3, respectively. In addition to providing a useful method for studying thiol protein kinetics, our studies add to recent reports challenging the popular belief that peroxiredoxins are poor enzymes toward hydrogen peroxide, as compared with heme and selenium proteins.
Keywords :
Peroxiredoxins , Peroxynitrite , Yeast , Antioxidant defense , Hydrogen peroxide-mediated signaling , hydrogen peroxide , Peroxiredoxin kinetics
Journal title :
Free Radical Biology and Medicine
Journal title :
Free Radical Biology and Medicine