Effect of thiocyanate on the peroxidase and pseudocatalase activities of Leishmania major ascorbate peroxidase
Metadata of CSIR Papers
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| Title |
Effect of thiocyanate on the peroxidase and pseudocatalase activities of Leishmania major ascorbate peroxidase
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| Creator |
Dolai, S
Yadav, RK Datta, AK Adak, S |
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| Subject |
Biochemistry & Molecular Biology; Biophysics
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| Description |
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 O-2. This pseudocatalatic degradation of H2O2 to O-2 is solely dependent oil ascorbate and is blocked by a spin trap, a-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 O-2 through the intermediate formation of O-2(-). 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 11 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 O-2. Binding studies by optical difference spectroscopy indicate that SCN- binds LmAPX (Kd 100 +/- 10 mM) near the herne edge. Thus, unlike mammalian peroxidases, SCN- acts as an inhibitor for Leishmania peroxidase to block ascorbate oxidation and subsequent pseudocatalase activity. (c) 2006 Elsevier B.V All rights reserved.
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| Publisher |
ELSEVIER SCIENCE BVAMSTERDAMPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
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| Date |
2011-09-20T12:12:41Z
2011-09-20T12:12:41Z 2007 |
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| Type |
Article
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| Identifier |
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
0304-4165 http://hdl.handle.net/123456789/14377 |
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| Language |
English
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