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High-field EPR and site-directed spin labeling reveal a periodical polarity profile: The sequence 88 to 94 of the phototransducer NpHtrII in complex with sensory rhodopsin, NpSRII

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Abstract

Taking advantage of the improved spectral resolution of high-field electron paramagnetic resonance (EPR) at 95 GHz/3.4 T as compared to conventional X-band EPR (9.5 GHz/0.34 T), detailed information on the polarity profile in a protein-protein interface is obtained. Nitroxide spin label side chains are introduced at positions 88 to 94 in the AS-1 sequence of the membrane adjacent HAMP domain of the transducer protein, NpHtrII, which is reconstituted in complex with sensory rhodopsin, NpSRII fromNatronobacterium pharaonis. Position-dependent variations of the values of the nitroxide magnetic tensor componentsg xx andA zz suggest that the spin label side chains at positions 88 to 93 of AS-1 are located between a hydrophobic and a hydrophilic microenvironment. The observed periodicity of the polarity properties of the respective spin label microenvironment agrees with an α-helical secondary structure of this part of AS-1 and validates a recently published molecular model which locates residues 88 and 91 in the interface between helices F and G of NpSRII and AS-1 of NpHtrII close to the cytoplasmic lipid-water interface.

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Authors and Affiliations

  1. Fachbereich Physik, Universität Osnabrück, Barbarastrasse 7, 49076, Osnabrück, Germany

    H. Brutlach, E. Bordignon, L. Urban, J. P. Klare, H. -J. Reyher & H. -J. Steinhoff

  2. Max-Planck-Institut für Molekulare Physiologie, Dortmund, Germany

    M. Engelhard

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  1. H. Brutlach
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  2. E. Bordignon
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  3. L. Urban
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  4. J. P. Klare
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  5. H. -J. Reyher
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  6. M. Engelhard
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  7. H. -J. Steinhoff
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Brutlach, H., Bordignon, E., Urban, L. et al. High-field EPR and site-directed spin labeling reveal a periodical polarity profile: The sequence 88 to 94 of the phototransducer NpHtrII in complex with sensory rhodopsin, NpSRII. Appl. Magn. Reson. 30, 359–372 (2006). https://doi.org/10.1007/BF03166206

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  • DOI: https://doi.org/10.1007/BF03166206

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