We demonstrate tracking of protein structural changes with time-resolved wide-angle X-ray scattering (TR-WAXS) with nanosecond time resolution. We investigated the tertiary and quaternary conformational changes of human hemoglobin under nearly physiological conditions triggered by laser-induced ligand photolysis. We also report data on optically induced tertiary relaxations of myoglobin and refolding of cytochrome c to illustrate the wide applicability of the technique. By providing insights into the structural dynamics of proteins functioning in their natural environment, TR-WAXS complements and extends results obtained with time-resolved optical spectroscopy and X-ray crystallography.
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We thank W.A. Eaton and E. Henry for helpful comments, H.-S. Cho and S. Ahn for helpful discussions about the data analysis, and Y.O. Jung, K.H. Kim and J.H. Lee for their assistance with sample preparation and experiments. This research was supported in part by the Intramural Research Program of the National Institutes of Health to P.A.A., by EU grant FLASH: FP6-503641 to M.W., and a grant from the Creative Research Initiatives (Center for Time-Resolved Diffraction) of the Ministry of Education, Science and Technology, Korea Science and Engineering Foundation to H.I.
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Cammarata, M., Levantino, M., Schotte, F. et al. Tracking the structural dynamics of proteins in solution using time-resolved wide-angle X-ray scattering. Nat Methods 5, 881–886 (2008). https://doi.org/10.1038/nmeth.1255
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