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Identifying conformational changes with site-directed spin labeling

Nature Structural Biology volume 7, pages 735–739 (2000)Cite this article

Abstract

Site-direct spin labeling combined with electron paramagnetic resonance (EPR) spectroscopy is a powerful tool for detecting structural changes in proteins. This review provides examples that illustrate strategies for interpreting the data in terms of specific rearrangements in secondary and tertiary structure. The changes in the mobility and solvent accessibility of the spin label side chains, and in the distances between spin labels, report (i) rigid body motions of α-helices and β-strands (ii) relative movements of domains and (iii) changes in secondary structure. Such events can be monitored in the millisecond timescale, making it possible to follow structural changes during function. There is no upper limit to the size of proteins that can be investigated, and only 50–100 picomoles of protein are required. These features make site-directed spin labeling an attractive approach for the study of structure and dynamics in a wide range of systems.

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Figure 1: The spin labeling reaction.
Figure 2: Effects of motion and magnetic dipolar interactions on nitroxide EPR spectra.
Figure 3: Light activation of rhodopsin.
Figure 4: Ribbon models of the open (blue) and closed (red) conformations of T4L.
Figure 5: Secondary structural change in a membrane transporter.

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References

  1. Hubbell, W.L. & Altenbach, C. Curr. Opin. Struct. Biol. 4, 566–573 (1994).

    Article  CAS  Google Scholar 

  2. Hubbell, W.L., Mchaourab, H.S., Altenbach, C., & Lietzow, M.A. Structure 4, 779–783 (1996).

    Article  CAS  PubMed Central  Google Scholar 

  3. Hubbell, W.L., Gross, A., Langen, R., & Lietzow, M.A. Curr. Opin. Struct. Biol. 8, 649–656 (1998).

    Article  CAS  Google Scholar 

  4. Feix, J.B. & Klug, C.S. In Biological Magnetic Resonance, Vol. 14: Spin Labeling: The Next Millenium (ed. Berliner, L.J.) 251–281 (Plenum Press, New York, New York; 1998).

    Google Scholar 

  5. Voss, J.C., Salwinski, L., Kaback, H.R. & Hubbell, W.L. Proc. Natl. Acad. Sci. USA 92, 12295–12299 (1995).

    Article  CAS  PubMed Central  Google Scholar 

  6. Oh, K.J., Altenbach, C., Collier, R.J. & Hubbell, W.L. Meth. Mol. Biol. 145, 147–169 (1999).

    Google Scholar 

  7. Griffith, O.H., Dehlinger, P.J. & Van. S.P. J. Membr. Biol. 15, 159–92 (1974).

    Article  CAS  PubMed Central  Google Scholar 

  8. Likhtensktein, G.I. Spin Labeling Methods in Molecular Biology. (Wiley, New York, New York; 1976.)

    Google Scholar 

  9. Rabenstein, M.D. & Shin, Y.-K. Proc. Natl. Acad. Sci. USA 92, 8239–8243 (1995).

    Article  CAS  PubMed Central  Google Scholar 

  10. Steinhoff, H.-J. et al. Biophys. J. 73, 3287–98. (1997).

    Article  CAS  PubMed Central  Google Scholar 

  11. Hustedt, E.J., Smirnov, A.I., Laub, C.F., Cobb, C.E. & Beth, A.H. Biophys. J. 74, 1861–1877 (1997).

    Article  Google Scholar 

  12. Hustedt, E.J. & Beth, A.H. Ann. Rev. Biophys. Biomol. Struct. 28, 129–153 (1999).

    Article  CAS  Google Scholar 

  13. Mchaourab, H.S., Oh, K.J., Fang, C.J. & Hubbell, W.L. Biochemistry 36, 307–316 (1997).

    Article  CAS  PubMed Central  Google Scholar 

  14. Langen, R., Oh, K.J., Cascio, D. & Hubbell, W.L. Biochemistry 39, in the press (2000).

  15. Koteiche, H.A. & Mchaourab, H.S. J. Mol. Biol. 294, 561–77 (1999).

    Article  CAS  PubMed Central  Google Scholar 

  16. Perozo, E., Cortes, D.M. & Cuello, L.G. Nature Struct. Biol. 5, 459–469 (1998).

    Article  CAS  PubMed Central  Google Scholar 

  17. Gross, A., Columbus, L., Hideg, K., Altenbach, C. & Hubbell, W.L. Biochemistry 38, 10324–10335 (1999).

    Article  CAS  Google Scholar 

  18. Poirier, M.A., Xiao, W., Macosko, J.C., Chan, C., Shin, Y.K. & Bennett, M.K. Nature Struct. Biol. 5, 765–9 (1998).

    Article  CAS  Google Scholar 

  19. Farrens, D., Altenbach, C., Yang, K., Hubbell, W.L. & Khorana, H.G. . Science 274, 768–770 (1996).

    Article  CAS  Google Scholar 

  20. Altenbach, C. et al. Biochemistry 35, 12470–12478 (1996).

    Article  CAS  Google Scholar 

  21. Lietzow, M.A. Ph.D. Thesis. Site-directed spin labeling of cellular retinol-binding protein. 1. Correlation of nitroxide side chain motion with protein structure and dynamics. 2. Analysis of ligand-induced conformational changes. (University of California, Los Angeles, California; 1999).

  22. Klug, C.S., Su, W. & Feix, J.B. Biochemistry 36, 13027–13033 (1997).

    Article  CAS  PubMed Central  Google Scholar 

  23. Perozo, E., Cortes, D.M. & Cuello, L.G. Science 285, 73–8 (1999).

    Article  CAS  Google Scholar 

  24. Ottemann, K.M., Xiao, W., Shin, Y.K., Koshland, D.E. Jr. Science 285, 1751–4. (1999).

    Article  CAS  PubMed Central  Google Scholar 

  25. Hall, J.A., Thorgeirsson, T.E., Liu, J., Shin, Y.K. & Nikaido, H. J. Biol. Chem. 272, 17610–17614 (1997).

    Article  CAS  PubMed Central  Google Scholar 

  26. Klug, C.S., Eaton, S.S., Eaton, G.R. & Feix, J.B. Biochemistry 37, 9016–9023 (1998).

    Article  CAS  PubMed Central  Google Scholar 

  27. Thorgeirsson, T.E. et al. J. Mol. Biol. 273, 951–957 (1997).

    Article  CAS  PubMed Central  Google Scholar 

  28. Rink, T., Pfeiffer, M., Oesterhelt, D., Gerwert, K. & Steinhoff, H.-J. Biophys. J. 78, 1519–1530 (2000).

    Article  CAS  PubMed Central  Google Scholar 

  29. Mollaaghababa, R., Steinhoff, H.-J., Hubbell, H.G. & Khorana, H.G. Biochemistry 39, 1120–1127 (2000).

    Article  CAS  PubMed Central  Google Scholar 

  30. Rink, T., Riesle, J., Oesterhelt, D., Gerwert, K. & Steinhoff, H.J. Biophys. J. 73, 983–93 (1997).

    Article  CAS  PubMed Central  Google Scholar 

  31. Steinhoff, H.J. et al. Science 266, 105–107 (1994).

    Article  CAS  PubMed Central  Google Scholar 

  32. Tiebel, B. et al. J. Mol. Biol. 290, 229–240 (1999).

    Article  CAS  PubMed Central  Google Scholar 

  33. Unger, V.M. & Schertler, G.F. Biophys. J. 68, 1776–1786 (1995).

    Article  CAS  PubMed Central  Google Scholar 

  34. Baldwin, J.M., Schertler, G.F. & Unger, V.M. J. Mol. Biol. 272, 144–64 (1997).

    Article  CAS  PubMed Central  Google Scholar 

  35. Farahbakhsh, Z., Ridge, K., Khorana, H.G. & Hubbell, W.L. Biochemistry 34, 8812–8819 (1995).

    Article  CAS  Google Scholar 

  36. Altenbach, C., Cai, K., Khorana, H.G. & Hubbell, W.L. Biochemistry 38, 7925–7930 (1999).

    Article  PubMed Central  Google Scholar 

  37. Altenbach, C., Klein-Seetharaman, J., Hwa, J., Khorana, H.G. & Hubbell, W.L. Biochemistry 38, 7945–7949 (1999).

    Article  CAS  PubMed Central  Google Scholar 

  38. Langen, R., Cai, K., Altenbach, C., Khorana, H.G. & Hubbell, W.L. Biochemistry 38, 7918–7924 (1999).

    Article  CAS  PubMed Central  Google Scholar 

  39. Mchaourab, H.S., Lietzow, M.A., Hideg, K. & Hubbell, W.L. Biochemistry 35, 7692–7704 (1996).

    Article  CAS  PubMed Central  Google Scholar 

  40. Farahbakhsh, Z., Hideg, K. & Hubbell, W.L. Science 262, 1416–1420 (1993).

    Article  CAS  PubMed Central  Google Scholar 

  41. Faber, H.R. & Matthews, B.W. Nature 348, 263–266 (1990).

    Article  CAS  Google Scholar 

  42. Mchaourab, H.S., Oh, K.J., Fang, C.J. & Hubbell, W.L. Biochemistry 35, 1404–14046 (1996).

    Article  Google Scholar 

  43. Merianos, H.J., Cadieus, N., Lin, C.H., Kadner, R.J. & Cafiso, D.S. Nature Struct. Biol. 7, 205–209 (2000).

    Article  CAS  Google Scholar 

  44. Kadner, R.J. Mol. Microbiol. 4, 2027–2033 (1990).

    Article  CAS  Google Scholar 

  45. Locher, K.P. et al. Cell 95, 771–778 (1998).

    Article  CAS  Google Scholar 

  46. Ferguson, A.D., Hofmann, E., Coulton, J.W., Diederichs, K. & Welte, W. Science 282, 2215–2220 (1998).

    Article  CAS  Google Scholar 

  47. Buchanan, S.K. et al. Nature Struct. Biol. 6, 56–63 (1999).

    Article  CAS  PubMed Central  Google Scholar 

  48. Jiang, X. et al. Science 276, 1261–1264 (1997).

    Article  CAS  Google Scholar 

  49. Palczewski, K. et al. Science 289, 739–745 (2000).

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Jules Stein Eye Institute and Department of Chemistry and Biochemistry, University of California, Los Angeles, 90095, California, USA

    Wayne L. Hubbell & Christian Altenbach

  2. Department of Chemistry, University of Virginia, Charlottesville, 22901, Virginia, USA

    David S. Cafiso

Authors
  1. Wayne L. Hubbell
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  2. David S. Cafiso
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  3. Christian Altenbach
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Correspondence to Wayne L. Hubbell.

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Hubbell, W., Cafiso, D. & Altenbach, C. Identifying conformational changes with site-directed spin labeling. Nat Struct Mol Biol 7, 735–739 (2000). https://doi.org/10.1038/78956

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