pH-Dependent structures of the manganese binding sites in oxalate decarboxylase as revealed by high-field electron paramagnetic resonance

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Tabares, Leandro C., Gätjens, Jessica, Hureau, Christelle, Burrell, Matthew R, Bowater, Laura, Pecoraro, Vincent L., Bornemann, Stephen and Un, Sun (2009) pH-Dependent structures of the manganese binding sites in oxalate decarboxylase as revealed by high-field electron paramagnetic resonance. The Journal of Physical Chemistry B, 113 (26). pp. 9016-9025. ISSN 1520-6106

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Abstract

A high-field electron paramagnetic resonance (HFEPR) study of oxalate decarboxylase (OxdC) is reported. OxdC breaks down oxalate to carbon dioxide and formate and possesses two distinct manganese(II) binding sites, referred to as site−1 and −2. The Mn(II) zero-field interaction was used to probe the electronic state of the metal ion and to examine chemical/mechanistic roles of each of the Mn(II) centers. High magnetic-fields were exploited not only to resolve the two sites, but also to measure accurately the Mn(II) zero-field parameters of each of the sites. The spectra exhibited surprisingly complex behavior as a function of pH. Six different species were identified based on their zero-field interactions, two corresponding to site-1 and four states to site-2. The assignments were verified using a mutant that only affected site-1. The speciation data determined from the HFEPR spectra for site −2 was consistent with a simple triprotic equilibrium model, while the pH dependence of site-1 could be described by a single pKa. This pH dependence was independent of the presence of the His-tag and of whether the preparations contained 1.2 or 1.6 Mn per subunit. Possible structures of the six species are proposed based on spectroscopic data from model complexes and existing protein crystallographic structures obtained at pH 8 are discussed. Although site-1 has been identified as the active site and no role has been assigned to site-2, the pronounced changes in the electronic structure of the latter and its pH behavior, which also matches the pH-dependent activity of this enzyme, suggests that even if the conversion of oxalate to formate is carried out at site-1, site-2 likely plays a catalytically relevant role.

Item Type:	Article
Faculty \ School:	Faculty of Medicine and Health Sciences > Norwich Medical School Faculty of Science > School of Biological Sciences Faculty of Science > School of Chemical Sciences and Pharmacy (former - to 2009)
Depositing User:	Rhiannon Harvey
Date Deposited:	31 May 2011 10:54
Last Modified:	12 Jan 2023 11:30
URI:	https://ueaeprints.uea.ac.uk/id/eprint/31554
DOI:	10.1021/jp9021807

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