Effect of thiocyanate on the peroxidase and pseudocatalase activities of Leishmania major ascorbate peroxidase

Dolai, Subhankar ; Yadav, Rajesh K. ; Datta, Alok K. ; Adak, Subrata (2007) Effect of thiocyanate on the peroxidase and pseudocatalase activities of Leishmania major ascorbate peroxidase Biochimica et Biophysica Acta (BBA) - General Subjects, 1770 (2). pp. 247-256. ISSN 0304-4165

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Official URL: http://linkinghub.elsevier.com/retrieve/pii/S03044...

Related URL: http://dx.doi.org/10.1016/j.bbagen.2006.10.001


We report here that the Leishmania major ascorbate peroxidase (LmAPX), having similarity with plant ascorbate peroxidase, catalyzes the oxidation of suboptimal concentration of ascorbate to monodehydroascorbate (MDA) at physiological pH in the presence of added H2O2 with concurrent evolution of O2. This pseudocatalatic degradation of H2O2 to O2 is solely dependent on ascorbate and is blocked by a spin trap, α-phenyl-n-tert-butyl nitrone (PBN), indicating the involvement of free radical species in the reaction process. LmAPX thus appears to catalyze ascorbate oxidation by its peroxidase activity, first generating MDA and H2O with subsequent regeneration of ascorbate by the reduction of MDA with H2O2 evolving O2 through the intermediate formation of O2-. Interestingly, both peroxidase and ascorbate-dependent pseudocatalatic activity of LmAPX are reversibly inhibited by SCN- in a concentration dependent manner. Spectral studies indicate that ascorbate cannot reduce LmAPX compound II to the native enzyme in presence of SCN-. Further kinetic studies indicate that SCN- itself is not oxidized by LmAPX but inhibits both ascorbate and guaiacol oxidation, which suggests that SCN- blocks initial peroxidase activity with ascorbate rather than subsequent nonenzymatic pseudocatalatic degradation of H2O2 to O2. Binding studies by optical difference spectroscopy indicate that SCN- binds LmAPX (Kd = 100 ± 10 mM) near the heme edge. Thus, unlike mammalian peroxidases, SCN- acts as an inhibitor for Leishmania peroxidase to block ascorbate oxidation and subsequent pseudocatalase activity.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:Leishmania; Ascorbate; Peroxidase; Thiocyanate; Pseudocatalase Activity
ID Code:9067
Deposited On:29 Oct 2010 11:45
Last Modified:30 May 2011 06:45

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