Characterization of Interactions Between Proteins Using Site-Directed Spin Labeling and Electron Paramagnetic Resonance Spectroscopy
Site-directed spin-labeling and the analysis of proteins by electron paramagnetic resonance spectroscopy provides a powerful tool for identifying sites of contact within protein complexes at the resolution of aminoacyl side chains. Here we describe the method as we have used it to study interactions of proteins involved in export via the Sec secretory system in Escherichia coli. The method is amendable to the study of most protein interactions.
Key wordsSite-directed spin labeling electron paramagnetic resonance spectroscopy EPR spin-labeled protein
- 8.Crane, J. M., Mao, C., Lilly, A. A., Smith, V. F., Suo, Y., Hubbell, W. L., and Randall, L. L. (2005) Mapping of the docking of SecA onto the chaperone SecB by site-directed spin labeling: insight into the mechanism of ligand transfer during protein export J. Mol. Biol. 353, 295–307.CrossRefPubMedGoogle Scholar
- 11.Fajer, P. G. (2000) in “Encyclopedia of Analytical Chemistry” (Meyers, R. A., Ed.), pp. 5725-61, John Wiley & Sons Ltd., London.Google Scholar
- 16.Schneider, D., and Freed, J. (1989) Spin Labeling: Theory and Application, Biological Magnetic Resonance (Berliner, L., and Reuben, J., Eds.), Vol. 8, Plenum, New York.Google Scholar