Oxidation of 4-tert-Butylcatechol and Dopamine by Hydrogen Peroxide Catalysed by Horseradisch Peroxidase
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M. García-Moreno
Abstract
The catalytic cycle of horseradish peroxidase (HRP; donor:hydrogen peroxide oxidoreductase; EC 1.11.1.7) is initiated by a rapid oxidation of it by hydrogen peroxide to give an enzyme intermediate, compound I, which reverts to the resting state via two successive single electron transfer reactions from reducing substrate molecules, the first yielding a second enzyme intermediate, compound II. To investigate the mechanism of action of horseradish peroxidase on catechol substrates we have studied the oxidation of both 4-tert-butylcatechol and dopamine catalysed by this enzyme. The different polarity of the side chains of both o-diphenol substrates could help in the understanding of the nature of the rate-limiting step in the oxidation of these substrates by the enzyme. The procedure used is based on the experimental data to the corresponding steady-state equations and permitted evaluation of the more significant individual rate constants involved in the corresponding reaction mechanism. The values obtained for the rate constants for each of the two substrates allow us to conclude that the reaction of horseradish peroxidase compound II with o-diphenols can be visualised as a two-step mechanism in which the first step corresponds to the formation of an enzyme-substrate complex, and the second to the electron transfer from the substrate to the iron atom. The size and hydrophobicity of the substrates control their access to the hydrophobic binding site of horseradish peroxidase, but electron density in the hydroxyl group of C-4 is the most important feature for the electron transfer step.
Copyright © 1999 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
- To Our Authors, Readers and Subscribers
- Michael Strauss. January 12, 1950 April 29, 1999
- Monoallelic Expression of Pax5: A Paradigm for the Haploinsufficiency of Mammalian Pax Genes?
- Adeno-Associated Viral Vectors for Gene Transfer and Gene Therapy
- Histidyl-tRNA Synthetase
- Dequalinium TM Vesicles Form Stable Complexes with Plasmid DNA which Are Protected from DNase Attack
- On the Role of Thymopoietins in Cell Proliferation. Immunochemical Evidence for New Members of the Human Thymopoietin Family
- Functional Characterization of Atherosclerosis-Associated Ser128Arg and Leu554Phe E-Selectin Mutations
- Cycloheximide, a New Tool to Dissect Specific Steps in ER-Associated Degradation of Different Substrates
- Localization of Rat Cathepsin K in Osteoclasts and Resorption Pits: Inhibition of Bone Resorption and Cathepsin K-Activity by Peptidyl Vinyl Sulfones
- Oxidation of 4-tert-Butylcatechol and Dopamine by Hydrogen Peroxide Catalysed by Horseradisch Peroxidase
- A Non-Radioactive Method for Inexpensive Quantitative RT-PCR
- Activation of the Transcription Factor ISGF3 by Interferon-gamma
- Acquisition of Myogenic Specificity through Replacement of One Amino Acid of MASH-1 and Introduction of an Additional alpha -Helical Turn
- The Heavy Metal-Responsive Transcription Factor-1 (MTF-1) Is Not Required for Neural Differentiation
- Human Fc gamma Receptor IIb Expressed in <I>Escherichia coli</I> Reveals IgG Binding Capability
- Mutational Analysis of Two Stefin A Epitopes
