RT Journal Article
SR Electronic
T1 Mapping Protein Dynamics at High-Resolution with Temperature-Jump X-ray Crystallography
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.06.10.495662
DO 10.1101/2022.06.10.495662
A1 Alexander M. Wolff
A1 Eriko Nango
A1 Iris D. Young
A1 Aaron S. Brewster
A1 Minoru Kubo
A1 Takashi Nomura
A1 Michihiro Sugahara
A1 Shigeki Owada
A1 Benjamin A. Barad
A1 Kazutaka Ito
A1 Asmit Bhowmick
A1 Sergio Carbajo
A1 Tomoya Hino
A1 James M. Holton
A1 Dohyun Im
A1 Lee J. O’Riordan
A1 Tomoyuki Tanaka
A1 Rie Tanaka
A1 Raymond G. Sierra
A1 Fumiaki Yumoto
A1 Kensuke Tono
A1 So Iwata
A1 Nicholas K. Sauter
A1 James S. Fraser
A1 Michael C. Thompson
YR 2022
UL http://biorxiv.org/content/early/2022/06/12/2022.06.10.495662.abstract
AB 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 trigger 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 sub-millisecond 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.Competing Interest StatementThe authors have declared no competing interest.