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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

  • H. Brutlach
  • E. Bordignon
  • L. Urban
  • J. P. Klare
  • H. -J. Reyher
  • M. Engelhard
  • H. -J. Steinhoff
Article

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.

Keywords

Electron Paramagnetic Resonance Electron Paramagnetic Resonance Spectrum Nitroxide Spin Label Polarity Profile 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer 2006

Authors and Affiliations

  • H. Brutlach
    • 1
  • E. Bordignon
    • 1
  • L. Urban
    • 1
  • J. P. Klare
    • 1
  • H. -J. Reyher
    • 1
  • M. Engelhard
    • 2
  • H. -J. Steinhoff
    • 1
  1. 1.Fachbereich PhysikUniversität OsnabrückOsnabrückGermany
  2. 2.Max-Planck-Institut für Molekulare PhysiologieDortmundGermany

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