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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References
Hubbell, W.L. & Altenbach, C. Curr. Opin. Struct. Biol. 4, 566–573 (1994).
Hubbell, W.L., Mchaourab, H.S., Altenbach, C., & Lietzow, M.A. Structure 4, 779–783 (1996).
Hubbell, W.L., Gross, A., Langen, R., & Lietzow, M.A. Curr. Opin. Struct. Biol. 8, 649–656 (1998).
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).
Voss, J.C., Salwinski, L., Kaback, H.R. & Hubbell, W.L. Proc. Natl. Acad. Sci. USA 92, 12295–12299 (1995).
Oh, K.J., Altenbach, C., Collier, R.J. & Hubbell, W.L. Meth. Mol. Biol. 145, 147–169 (1999).
Griffith, O.H., Dehlinger, P.J. & Van. S.P. J. Membr. Biol. 15, 159–92 (1974).
Likhtensktein, G.I. Spin Labeling Methods in Molecular Biology. (Wiley, New York, New York; 1976.)
Rabenstein, M.D. & Shin, Y.-K. Proc. Natl. Acad. Sci. USA 92, 8239–8243 (1995).
Steinhoff, H.-J. et al. Biophys. J. 73, 3287–98. (1997).
Hustedt, E.J., Smirnov, A.I., Laub, C.F., Cobb, C.E. & Beth, A.H. Biophys. J. 74, 1861–1877 (1997).
Hustedt, E.J. & Beth, A.H. Ann. Rev. Biophys. Biomol. Struct. 28, 129–153 (1999).
Mchaourab, H.S., Oh, K.J., Fang, C.J. & Hubbell, W.L. Biochemistry 36, 307–316 (1997).
Langen, R., Oh, K.J., Cascio, D. & Hubbell, W.L. Biochemistry 39, in the press (2000).
Koteiche, H.A. & Mchaourab, H.S. J. Mol. Biol. 294, 561–77 (1999).
Perozo, E., Cortes, D.M. & Cuello, L.G. Nature Struct. Biol. 5, 459–469 (1998).
Gross, A., Columbus, L., Hideg, K., Altenbach, C. & Hubbell, W.L. Biochemistry 38, 10324–10335 (1999).
Poirier, M.A., Xiao, W., Macosko, J.C., Chan, C., Shin, Y.K. & Bennett, M.K. Nature Struct. Biol. 5, 765–9 (1998).
Farrens, D., Altenbach, C., Yang, K., Hubbell, W.L. & Khorana, H.G. . Science 274, 768–770 (1996).
Altenbach, C. et al. Biochemistry 35, 12470–12478 (1996).
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).
Klug, C.S., Su, W. & Feix, J.B. Biochemistry 36, 13027–13033 (1997).
Perozo, E., Cortes, D.M. & Cuello, L.G. Science 285, 73–8 (1999).
Ottemann, K.M., Xiao, W., Shin, Y.K., Koshland, D.E. Jr. Science 285, 1751–4. (1999).
Hall, J.A., Thorgeirsson, T.E., Liu, J., Shin, Y.K. & Nikaido, H. J. Biol. Chem. 272, 17610–17614 (1997).
Klug, C.S., Eaton, S.S., Eaton, G.R. & Feix, J.B. Biochemistry 37, 9016–9023 (1998).
Thorgeirsson, T.E. et al. J. Mol. Biol. 273, 951–957 (1997).
Rink, T., Pfeiffer, M., Oesterhelt, D., Gerwert, K. & Steinhoff, H.-J. Biophys. J. 78, 1519–1530 (2000).
Mollaaghababa, R., Steinhoff, H.-J., Hubbell, H.G. & Khorana, H.G. Biochemistry 39, 1120–1127 (2000).
Rink, T., Riesle, J., Oesterhelt, D., Gerwert, K. & Steinhoff, H.J. Biophys. J. 73, 983–93 (1997).
Steinhoff, H.J. et al. Science 266, 105–107 (1994).
Tiebel, B. et al. J. Mol. Biol. 290, 229–240 (1999).
Unger, V.M. & Schertler, G.F. Biophys. J. 68, 1776–1786 (1995).
Baldwin, J.M., Schertler, G.F. & Unger, V.M. J. Mol. Biol. 272, 144–64 (1997).
Farahbakhsh, Z., Ridge, K., Khorana, H.G. & Hubbell, W.L. Biochemistry 34, 8812–8819 (1995).
Altenbach, C., Cai, K., Khorana, H.G. & Hubbell, W.L. Biochemistry 38, 7925–7930 (1999).
Altenbach, C., Klein-Seetharaman, J., Hwa, J., Khorana, H.G. & Hubbell, W.L. Biochemistry 38, 7945–7949 (1999).
Langen, R., Cai, K., Altenbach, C., Khorana, H.G. & Hubbell, W.L. Biochemistry 38, 7918–7924 (1999).
Mchaourab, H.S., Lietzow, M.A., Hideg, K. & Hubbell, W.L. Biochemistry 35, 7692–7704 (1996).
Farahbakhsh, Z., Hideg, K. & Hubbell, W.L. Science 262, 1416–1420 (1993).
Faber, H.R. & Matthews, B.W. Nature 348, 263–266 (1990).
Mchaourab, H.S., Oh, K.J., Fang, C.J. & Hubbell, W.L. Biochemistry 35, 1404–14046 (1996).
Merianos, H.J., Cadieus, N., Lin, C.H., Kadner, R.J. & Cafiso, D.S. Nature Struct. Biol. 7, 205–209 (2000).
Kadner, R.J. Mol. Microbiol. 4, 2027–2033 (1990).
Locher, K.P. et al. Cell 95, 771–778 (1998).
Ferguson, A.D., Hofmann, E., Coulton, J.W., Diederichs, K. & Welte, W. Science 282, 2215–2220 (1998).
Buchanan, S.K. et al. Nature Struct. Biol. 6, 56–63 (1999).
Jiang, X. et al. Science 276, 1261–1264 (1997).
Palczewski, K. et al. Science 289, 739–745 (2000).
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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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DOI: https://doi.org/10.1038/78956
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