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Site-directed spin labeling EPR spectroscopy in protein research

  • Johann P. Klare

    Johann P. Klare studied Chemistry in Dortmund and received his PhD in 2003 at the Max Planck Institute for Molecular Physiology, Dortmund. Until 2006, he was a Postdoc at the Max Planck Institute, studying the structural and mechanistic basis underlying signal transduction in archaeal phototaxis complexes. In 2006, he joined the group of Heinz-Jürgen Steinhoff at the University of Osnabrück. His research group applies EPR spectroscopy and other biophysical methods to study the structure, function and conformational dynamics of proteins, protein-protein and protein-nucleic acid complexes with a focus on biomedical applications.

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Published/Copyright: August 2, 2013

Abstract

Site-directed spin labeling (SDSL) in combination with electron paramagnetic resonance (EPR) spectroscopy has emerged as an efficient tool to elucidate the structure and the conformational dynamics of proteins under conditions close to the native state. This review article summarizes the basics as well as the recent progress in SDSL and EPR methods, especially for investigations on protein structure, protein function, and interaction of proteins with other proteins or nucleic acids. Labeling techniques as well as EPR methods are introduced and exemplified with applications to systems that have been studied in the author’s laboratory in the past 15 years, headmost the sensory rhodopsin-transducer complex mediating the photophobic response of the halophilic archaeum Natronomonas pharaonis. Further examples underline the application of SDSL EPR spectroscopy to answer specific questions about the system under investigation, such as the nature and influence of interactions of proteins with other proteins or nucleic acids. Finally, it is discussed how SDSL EPR can be combined with other biophysical techniques to combine the strengths of the different methodologies.


Corresponding author: Johann P. Klare, Physics Department, University of Osnabrück, Barbarastr. 7, D-49076 Osnabrück, Germany

About the author

Johann P. Klare

Johann P. Klare studied Chemistry in Dortmund and received his PhD in 2003 at the Max Planck Institute for Molecular Physiology, Dortmund. Until 2006, he was a Postdoc at the Max Planck Institute, studying the structural and mechanistic basis underlying signal transduction in archaeal phototaxis complexes. In 2006, he joined the group of Heinz-Jürgen Steinhoff at the University of Osnabrück. His research group applies EPR spectroscopy and other biophysical methods to study the structure, function and conformational dynamics of proteins, protein-protein and protein-nucleic acid complexes with a focus on biomedical applications.

I gratefully acknowledge K.F. Pirker for the careful reading of the article and helpful suggestions and comments.

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Received: 2013-4-2
Accepted: 2013-7-31
Published Online: 2013-08-02
Published in Print: 2013-10-01

©2013 by Walter de Gruyter Berlin Boston

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