Luque, DanielOrtega-Esteban, AlvaroValbuena, AlejandroVilas, Jose LuisRodríguez-Huete, AliciaMateu, Mauricio GCastón, José R2024-01-162024-01-162023-04-15J Mol Biol. 2023 Apr 15;435(8):168024.http://hdl.handle.net/20.500.12105/17180The biological function of macromolecular complexes depends not only on large-scale transitions between conformations, but also on small-scale conformational fluctuations at equilibrium. Information on the equilibrium dynamics of biomolecular complexes could, in principle, be obtained from local resolution (LR) data in cryo-electron microscopy (cryo-EM) maps. However, this possibility had not been validated by comparing, for a same biomolecular complex, LR data with quantitative information on equilibrium dynamics obtained by an established solution technique. In this study we determined the cryo-EM structure of the minute virus of mice (MVM) capsid as a model biomolecular complex. The LR values obtained correlated with crystallographic B factors and with hydrogen/deuterium exchange (HDX) rates obtained by mass spectrometry (HDX-MS), a gold standard for determining equilibrium dynamics in solution. This result validated a LR-based cryo-EM approach to investigate, with high spatial resolution, the equilibrium dynamics of biomolecular complexes. As an application of this approach, we determined the cryo-EM structure of two mutant MVM capsids and compared their equilibrium dynamics with that of the wild-type MVM capsid. The results supported a previously suggested linkage between mechanical stiffening and impaired equilibrium dynamics of a virus particle. Cryo-EM is emerging as a powerful approach for simultaneously acquiring information on the atomic structure and local equilibrium dynamics of biomolecular complexes.engVoRhttp://creativecommons.org/licenses/by-nc-nd/4.0/Equilibrium dynamicsMechanical elasticityCryo-electron microscopyAtomic force microscopyVirus capsidVirus capsidAttribution-NonCommercial-NoDerivatives 4.0 Internacional36828271435816802410.1016/j.jmb.2023.1680241089-8638Journal of molecular biologyopen access