RT Journal Article
SR Electronic
T1 3D Variability Analysis: Resolving continuous flexibility and discrete heterogeneity from single particle cryo-EM
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.04.08.032466
DO 10.1101/2020.04.08.032466
A1 Ali Punjani
A1 David J. Fleet
YR 2021
UL http://biorxiv.org/content/early/2021/01/13/2020.04.08.032466.abstract
AB Single particle cryo-EM excels in determining static structures of protein molecules, but existing 3D reconstruction methods have been ineffective in modelling flexible proteins. We introduce 3D variability analysis (3DVA), an algorithm that fits a linear subspace model of conformational change to cryo-EM data at high resolution. 3DVA enables the resolution and visualization of detailed molecular motions of both large and small proteins, revealing new biological insight from single particle cryo-EM data. Experimental results demonstrate the ability of 3DVA to resolve multiple flexible motions of α-helices in the sub-50 kDa transmembrane domain of a GPCR complex, bending modes of a sodium ion channel, five types of symmetric and symmetry-breaking flexibility in a proteasome, large motions in a spliceosome complex, and discrete conformational states of a ribosome assembly. 3DVA is implemented in the cryoSPARC software package.Competing Interest StatementA.P. is CEO of Stuctura Biotechnology Inc. which builds the cryoSPARC software package, distributed freely for academic non-profit use. D.J.F. is an advisor to Stuctura Biotechnology Inc. The novel aspects of the method presented are described in a provisional patent application.