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Towards an identification of chemically different flavin radicals by means of theirg-tensor

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Abstract

Theg-tensors of two chemically different flavin mononucleotide (FMN) radicals, one of which is covalently bound via N(5) of its 7,8-dimethyl isoalloxazine moiety, and the other one non-covalently bound to mutant LOV domains of the blue-light receptor phototropin, LOV1 C57M and LOV2 C450A, respectively, have been determined by very high microwave frequency and high magnetic field electron paramagnetic resonance (EPR) performed at 360 GHz and 12.8 T. Due to the high spectral resolution of the frozen-solution continuous-wave EPR spectra, the anisotropy of theg-tensors could be fully resolved. By least-squares fittings of spectral simulations to expermental data, the principal values ofg have been established:g X=2.00554(5),g Y=2.00391(5), andg Z=2.00247(7) for the N(5)-alkyl-chain-linked FMN radical in LOV1 C57M-675, andg X=2.00427(5),g Y=2.00360(5), andg Z=2.00220(7) for the noncovalently bound FMN radical in LOV2 C450A-605. By a comparison of these values to the ones from the flavin adenine dinucleotide radicals in two photolyases, the radical in LOV2 C450A-605 could be clearly identified as a neutral FMN radical, FMNH. In contrast, LOV1 C57M-675 exhibits significantly shifted principal components ofg, the differences being caused by spin-orbit coupling of the nearby sulfur from the reactive methionine residue, and the modified chemical structure due to the covalent attachment at N(5) of the radical to the apoprotein. The results clearly show the potential of using theg-tensor as probe of the global electronic and chemical structure of protein-bound flavin radicals.

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Author information

Author notes

  1. M. Fischer

    Present address: Institut für Biochemie und Lebensmittelchemie, Universität Hamburg, Hamburg, Germany

  2. M. R. Fuchs

    Present address: Institut für Kristallographie c/o BESSY, Freie Universität Berlin, Berlin, Germany

  3. C. W. M. Kay

    Present address: Department of Biology, University College London, London, UK

  4. G. Richter

    Present address: School of Chemistry, Cardiff University, Cardiff, UK

Authors and Affiliations

  1. Fachbereich Physik, Freie Universität Berlin, Arnimallee 14, 14195, Berlin, Germany

    A. Schnegg, A. Okafuji, R. Bittl, M. R. Fuchs, C. W. M. Kay, E. Schleicher & S. Weber

  2. Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Garching, Germany

    A. Bacher, M. Fischer, M. Joshi & G. Richter

  3. Institut für Biologie und Experimentelle Biophysik, Humboldt-Universität zu Berlin, Berlin, Germany

    P. Hegemann

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Schnegg, A., Okafuji, A., Bacher, A. et al. Towards an identification of chemically different flavin radicals by means of theirg-tensor. Appl. Magn. Reson. 30, 345–358 (2006). https://doi.org/10.1007/BF03166205

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  • DOI: https://doi.org/10.1007/BF03166205

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