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Exploring the Fe(III) binding sites of human serum transferrin with EPR at 275 GHz

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Abstract

We report 275 GHz EPR spectra of human serum transferrin. At this high microwave frequency the zero-field splitting between the magnetic sublevels of the high-spin \(\text{Fe}^{3+}\) sites can be accurately determined. We find the zero-field splitting to be a sensitive probe of the structure of the transferrin iron-binding sites. Signals arising from iron bound to the transferrin N-lobe can clearly be distinguished from signals from iron bound to the C-lobe. Moreover, our spectra show that the structure of the iron site in the N-lobe is influenced by the presence and conformation of the C-lobe. The spectra of a series of N-lobe mutants altering the second-shell interaction of Arg124 with the synergistic anion carbonate reflect conformational changes induced at the iron site.

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Acknowledgments

The research was supported with financial aid by the Netherlands Organization for Scientific Research (NWO), Department of Chemical Sciences (CW). GM gratefully acknowledges the Rubicon Fellowship from the Netherlands Organization for Scientific Research.

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  1. Guinevere Mathies

    Present address: Department of Chemistry, Francis Bitter Magnet Laboratory, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA

Authors and Affiliations

  1. Department of Physics, Huygens-Kamerlingh Onnes Laboratory, Leiden University, Leiden, The Netherlands

    Guinevere Mathies, Peter Gast & Edgar J. J. Groenen

  2. Department of Chemistry, Parsons Hall, University of New Hampshire, Durham, NH, 03824, USA

    N. Dennis Chasteen

  3. Department of Biochemistry, University of Vermont College of Medicine, Burlington, VT, 05405, USA

    Ashley N. Luck & Anne B. Mason

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  1. Guinevere Mathies
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Correspondence to Guinevere Mathies.

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Mathies, G., Gast, P., Chasteen, N.D. et al. Exploring the Fe(III) binding sites of human serum transferrin with EPR at 275 GHz. J Biol Inorg Chem 20, 487–496 (2015). https://doi.org/10.1007/s00775-014-1229-z

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  • DOI: https://doi.org/10.1007/s00775-014-1229-z

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