Accessibility navigation


Thermodynamic analysis of ferrous ion binding to Escherichia coli ferritin EcFtnA

Bou-Abdallah, F., Woodhall, M. R., Velazquez-Campoy, A., Andrews, S. C. and Chasteen, N. D. (2005) Thermodynamic analysis of ferrous ion binding to Escherichia coli ferritin EcFtnA. Biochemistry, 44 (42). pp. 13837-13846. ISSN 0006-2960

Full text not archived in this repository.

To link to this article DOI: 10.1021/bi0514212

Abstract/Summary

Iron oxidation in the bacterial ferritin EcFtnA from Escherichia coli shows marked differences from its homologue human H-chain ferritin (HuHF). While the amino acid residues that constitute the dinuclear center in these proteins are highly conserved, EcFtnA has a third iron-binding site (C site) in close proximity to the dinuclear center that is seemingly responsible for these differences. Here, we describe the first thermodynamic study of Fe2+ binding to EcFtnA and its variants to determine the location of the primary ferrous ion-binding sites on the protein and to better understand the role of the third C site in iron binding. Isothermal titration calorimetric analyses of the wild-type protein reveal the presence of two main classes of binding sites in the pH range of 6.5-7.5, ascribed to Fe2+ binding, first at the A and then the B sites. Site-directed mutagenesis of ligands in the A, B, or C sites affects the apparent Fe2+-binding stoichiometries at the unaltered sites. The data imply some degree of inter- and intrasubunit negative cooperative interaction between sites. Unlike HuHF where only the A site initially binds Fe2+, both A and B sites in EcFtnA bind Fe2+, implying a role for the C site in influencing the binding of Fe2+ at the B site of the di-iron center of EcFtnA. The ITC equations describing a binding model for three classes of independent binding sites are reported here for the first time.

Item Type:Article
Refereed:Yes
Divisions:Faculty of Life Sciences > School of Biological Sciences
ID Code:10286
Uncontrolled Keywords:ISOTHERMAL TITRATION CALORIMETRY, H-CHAIN FERRITIN, IRON STORAGE, FUNCTION, BOVINE SERUM-ALBUMIN, METAL-BINDING, OXIDATION, BACTERIOFERRITIN, DETOXIFICATION, CHEMISTRY, BACTERIA

University Staff: Request a correction | Centaur Editors: Update this record

Page navigation