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Evaluation of Conformational dynamics properties of spin-labelled proteins at different degrees of nitroxide radicals immobilization

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Abstract

Spin-labelling has found wide applications in elucidation of the dynamic behaviour of biological macromolecules in aqueous media and biomembranes. Most of the proposed methods aimed at estimation of macromolecular correlation times (τc) assume, however, spin label molecules rigidly bound within the protein matrix. To avoid this limitation theoretical models which involve additional dynamic parameters to characterize the spin label motion should be considered. We have used ESR spectra analysis technique which permits quantitative separation of slow macromolecular rotation (described by the rotational correlation time, τc) and fast anisotropic relative to protein nitroxyl radical motion (described by the “order parameter”,S). This method was applied to study: i) conformational dynamics of covalently and non-covalently spin-labelled human serum albumin (HSA) in solution; ii) protein-protein (antigen-antibody) interactions in a model system containing spin-labelled bovine serum albumin (BSA) and anti-BSA immunoglobulin (IgG) in solution; and iii) dynamic properties of membrane-bound proteins: H+-ATPase (CF1-CF0 coupling factor of photophosphorylation) and Photosystem I pigment-protein reaction centre complex (PSI RC) isolated from spinach chloroplasts and reconstituted in proteoliposomes.

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Author notes

  1. T. G. Gantchev

    Present address: Medical Research Council Group in the Radiation Sciences, Faculty of Medicine, University of Sherbrooke, J1H 5N4, Sherbrooke, Quebec, Canada

Authors and Affiliations

  1. Research Group in Cellular and Molecular Biology of Cancer; Oncochromosome Laboratory, National Centre of Oncology, Bulgarian Academy of Medicine, 1156, Sofia, Bulgaria

    T. G. Gantchev

  2. Faculty of Chemistry, Sofia University, Sofia, Bulgaria

    G. G. Gotchev

Authors
  1. T. G. Gantchev
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  2. G. G. Gotchev
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Gantchev, T.G., Gotchev, G.G. Evaluation of Conformational dynamics properties of spin-labelled proteins at different degrees of nitroxide radicals immobilization. Appl. Magn. Reson. 3, 67–82 (1992). https://doi.org/10.1007/BF03166781

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  • DOI: https://doi.org/10.1007/BF03166781

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