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Site-Directed Spin Labeling of Membrane Proteins

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Membrane Protein Structure

Part of the book series: Methods in Physiology Series ((METHPHYS))

Abstract

Membrane-associated proteins present a particularly difficult problem for structure determination. Diffraction approaches are hampered by the lack of crystallization methods, and multidimensional nuclear magnetic resonance (NMR)-approaches are not generally applicable due to the lack of high-resolution spectra (but see Opella, Chapter 11). For this class of proteins, the technique of spin labeling offers an attractive alternative for obtaining structural information. In this method, a stable nitroxide-free radical (the “spin label”) is attached at a specific site in the system of interest, and the electron paramagnetic resonance (EPR) spectrum is analyzed to yield information regarding the local environment around the label. Humphries and McConnell (1982) and Marsh (1981) have written excellent reviews of the technique and its capabilities. The two-volume series, Spin Labeling: Theory and Application, (Berliner, 1976, 1979) and Volume 8 of the series, Biological Magnetic Resonance, (Berliner and Reuben, 1990) provide a comprehensive treatment of continuous-wave EPR aspects of the subject. Two recent books, EPR and Advanced EPR Studies of Biological Systems (Dalton, 1985) and Advanced EPR: Applications in Biology and Biochemistry (Hoff, 1989), treat more advanced concepts and time-domain EPR.

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Authors
  1. Wayne L. Hubbell
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  2. Christian Altenbach
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Editors and Affiliations

  1. College of Medicine, Department of Physiology and Biophysics, University of California, Irvine, Irvine, California, USA

    Stephen H. White Ph.D. (Professor of Physiology and Biophysics) (Professor of Physiology and Biophysics)

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© 1994 American Physiological Society

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Hubbell, W.L., Altenbach, C. (1994). Site-Directed Spin Labeling of Membrane Proteins. In: White, S.H. (eds) Membrane Protein Structure. Methods in Physiology Series. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-7515-6_10

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  • DOI: https://doi.org/10.1007/978-1-4614-7515-6_10

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