Univ. of California, Merced, CA (United States); SLAC
RIKEN SPring-8 Center, Sayo (Japan); Tohoku Univ., Sendai (Japan)
Univ. of California, San Francisco, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
RIKEN SPring-8 Center, Sayo (Japan); Univ. of Hyogo (Japan)
RIKEN SPring-8 Center, Sayo (Japan)
The Scripps Research Inst., La Jolla, CA (United States); Univ. of California, San Francisco, CA (United States)
Asahi Kasei Pharma Corporation, Izunokuni (Japan)
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States); Univ. of California, Los Angeles, CA (United States)
Tottori Univ. (Japan)
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Univ. of California, San Francisco, CA (United States); SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Kyoto Univ. (Japan)
RIKEN SPring-8 Center, Sayo (Japan); Kyoto Univ. (Japan)
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Ginward Japan K.K., Tokyo (Japan); Inst. of Materials Structure Science (IMSS), Tsukuba, Japan
RIKEN SPring-8 Center, Sayo (Japan); Japan Synchrotron Radiation Research Institute, Sayo, Hyogo (Japan)
Univ. of California, San Francisco, CA (United States)
Understanding and controlling protein motion at atomic resolution is a hallmark challenge for structural biologists and protein engineers because conformational dynamics are essential for complex functions such as enzyme catalysis and allosteric regulation. Time-resolved crystallography offers a window into protein motions, yet without a universal perturbation to initiate conformational changes the method has been limited in scope. Here we couple a solvent-based temperature jump with time-resolved crystallography to visualize structural motions in lysozyme, a dynamic enzyme. We observed widespread atomic vibrations on the nanosecond timescale, which evolve on the submillisecond timescale into localized structural fluctuations that are coupled to the active site. An orthogonal perturbation to the enzyme, inhibitor binding, altered these dynamics by blocking key motions that allow energy to dissipate from vibrations into functional movements linked to the catalytic cycle. Because temperature jump is a universal method for perturbing molecular motion, the method demonstrated here is broadly applicable for studying protein dynamics.
Wolff, Alexander M., et al. "Mapping protein dynamics at high spatial resolution with temperature-jump X-ray crystallography." Nature Chemistry, vol. 15, no. 11, Sep. 2023. https://doi.org/10.1038/s41557-023-01329-4
Wolff, Alexander M., Nango, Eriko, Young, Iris D., Brewster, Aaron S., Kubo, Minoru, Nomura, Takashi, Sugahara, Michihiro, Owada, Shigeki, Barad, Benjamin A., Ito, Kazutaka, Bhowmick, Asmit, Carbajo, Sergio, Hino, Tomoya, Holton, James M., Im, Dohyun, O’Riordan, Lee J., Tanaka, Tomoyuki, Tanaka, Rie, ... Thompson, Michael C. (2023). Mapping protein dynamics at high spatial resolution with temperature-jump X-ray crystallography. Nature Chemistry, 15(11). https://doi.org/10.1038/s41557-023-01329-4
Wolff, Alexander M., Nango, Eriko, Young, Iris D., et al., "Mapping protein dynamics at high spatial resolution with temperature-jump X-ray crystallography," Nature Chemistry 15, no. 11 (2023), https://doi.org/10.1038/s41557-023-01329-4
@article{osti_2405208,
author = {Wolff, Alexander M. and Nango, Eriko and Young, Iris D. and Brewster, Aaron S. and Kubo, Minoru and Nomura, Takashi and Sugahara, Michihiro and Owada, Shigeki and Barad, Benjamin A. and Ito, Kazutaka and others},
title = {Mapping protein dynamics at high spatial resolution with temperature-jump X-ray crystallography},
annote = {Understanding and controlling protein motion at atomic resolution is a hallmark challenge for structural biologists and protein engineers because conformational dynamics are essential for complex functions such as enzyme catalysis and allosteric regulation. Time-resolved crystallography offers a window into protein motions, yet without a universal perturbation to initiate conformational changes the method has been limited in scope. Here we couple a solvent-based temperature jump with time-resolved crystallography to visualize structural motions in lysozyme, a dynamic enzyme. We observed widespread atomic vibrations on the nanosecond timescale, which evolve on the submillisecond timescale into localized structural fluctuations that are coupled to the active site. An orthogonal perturbation to the enzyme, inhibitor binding, altered these dynamics by blocking key motions that allow energy to dissipate from vibrations into functional movements linked to the catalytic cycle. Because temperature jump is a universal method for perturbing molecular motion, the method demonstrated here is broadly applicable for studying protein dynamics.},
doi = {10.1038/s41557-023-01329-4},
url = {https://www.osti.gov/biblio/2405208},
journal = {Nature Chemistry},
issn = {ISSN 1755-4330},
number = {11},
volume = {15},
place = {United States},
publisher = {Nature Publishing Group},
year = {2023},
month = {09}}
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC); National Science Foundation (NSF); National Institutes of Health (NIH); Japan Agency for Medical Research and Development (AMED); Japan Synchrotron Radiation Research Institute (JASRI); Japan Society for the Promotion of Science (JSPS)
Coherent X-ray Imaging Data Bank (Lawrence Berkeley National Laboratory); The University of Tokyo, RIKEN SPring-8 Centerhttps://doi.org/10.11577/1376050
