PMID:14673713

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Citation

Aitken-Rogers, H, Singleton, C, Lewin, A, Taylor-Gee, A, Moore, GR and Le Brun, NE (2004) Effect of phosphate on bacterioferritin-catalysed iron(II) oxidation. J. Biol. Inorg. Chem. 9:161-70

Abstract

The iron(III) mineral cores of bacterioferritins (BFRs), as isolated, contain a significant component of phosphate, with an iron-to-phosphate ratio approaching 1:1 in some cases. In order to better understand the in vivo core-formation process, the effect of phosphate on in vitro core formation in Escherichia coli BFR was investigated. Iron cores reconstituted in the presence of phosphate were found to have iron-to-phosphate ratios similar to those of native cores, and possessed electron paramagnetic resonance properties characteristic of the phosphate-rich core. Phosphate did not affect the stoichiometry of the initial iron(II) oxidation reaction that takes place at the intrasubunit dinuclear iron-binding sites (phase 2 of core formation), but did increase the rate of oxidation. Phosphate had a more significant effect on subsequent core formation (the phase 3 reaction), increasing the rate up to five-fold at pH 6.5 and 25 degrees C. The dependence of the phase 3 rate on phosphate was complex, being greatest at low phosphate and gradually decreasing until the point of saturation at approximately 2 mM phosphate (for iron(II) concentrations <200 microM). Phosphate caused a significant decrease in the absorption properties of both phase 2 and phase 3 products, and the phosphate dependence of the latter mirrored the observed rate dependence, suggesting that distinct iron(III)-phosphate species are formed at different phosphate concentrations. The effect of phosphate on absorption properties enabled the observation of previously undetected events in the phase 2 to phase 3 transition period.

Links

PubMed Online version:10.1007/s00775-003-0504-1

Keywords

Bacterial Proteins/chemistry; Catalysis; Chromatography, Ion Exchange; Cytochrome b Group/chemistry; Ferritins/chemistry; Ferrous Compounds/chemistry; Hydrogen-Ion Concentration; Kinetics; Models, Molecular; Oxidation-Reduction; Phosphates/chemistry; Protein Conformation

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