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Structural and spectroscopic characterisation of a heme peroxidase from sorghum Topical Issue in Honor of R.J.P. Williams

Nnamchi, CI and Parkin, G and Efimov, I and Basran, J and Kwon, H and Svistunenko, DA and Agirre, J and Okolo, BN and Moneke, A and Nwanguma, BC and Moody, PCE and Raven, EL (2016) 'Structural and spectroscopic characterisation of a heme peroxidase from sorghum Topical Issue in Honor of R.J.P. Williams.' Journal of Biological Inorganic Chemistry, 21 (1). 63 - 70. ISSN 0949-8257

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Abstract

A cationic class III peroxidase from Sorghum bicolor was purified to homogeneity. The enzyme contains a high-spin heme, as evidenced by UV-visible spectroscopy and EPR. Steady state oxidation of guaiacol was demonstrated and the enzyme was shown to have higher activity in the presence of calcium ions. A Fe /Fe reduction potential of-266 mV vs NHE was determined. Stopped-flow experiments with H O showed formation of a typical peroxidase Compound I species, which converts to Compound II in the presence of calcium. A crystal structure of the enzyme is reported, the first for a sorghum peroxidase. The structure reveals an active site that is analogous to those for other class I heme peroxidase, and a substrate binding site (assigned as arising from binding of indole-3-acetic acid) at the γ-heme edge. Metal binding sites are observed in the structure on the distal (assigned as a Na ion) and proximal (assigned as a Ca ) sides of the heme, which is consistent with the Ca -dependence of the steady state and pre-steady state kinetics. It is probably the case that the structural integrity (and, thus, the catalytic activity) of the sorghum enzyme is dependent on metal ion incorporation at these positions. III II + 2+ 2+ 2 2

Item Type: Article
Subjects: Q Science > QD Chemistry
Divisions: Faculty of Science and Health > Life Sciences, School of
Depositing User: Jim Jamieson
Date Deposited: 07 Apr 2016 12:01
Last Modified: 07 Apr 2021 10:16
URI: http://repository.essex.ac.uk/id/eprint/16382

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