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


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.


Electron Paramagnetic Resonance Electron Paramagnetic Resonance Spectrum Nitroxide Spin Label Polarity Profile 
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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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