@article{aebbe660ef9e4d7ab76c667860ee8325,
title = "Separating distinct structures of multiple macromolecular assemblies from cryo-EM projections",
abstract = "Single particle analysis for structure determination in cryo-electron microscopy is traditionally applied to samples purified to near homogeneity as current reconstruction algorithms are not designed to handle heterogeneous mixtures of structures from many distinct macromolecular complexes. We extend on long established methods and demonstrate that relating two-dimensional projection images by their common lines in a graphical framework is sufficient for partitioning distinct protein and multiprotein complexes within the same data set. The feasibility of this approach is first demonstrated on a large set of synthetic reprojections from 35 unique macromolecular structures spanning a mass range of hundreds to thousands of kilodaltons. We then apply our algorithm on cryo-EM data collected from a mixture of five protein complexes and use existing methods to solve multiple three-dimensional structures ab initio. Incorporating methods to sort single particle cryo-EM data from extremely heterogeneous mixtures will alleviate the need for stringent purification and pave the way toward investigation of samples containing many unique structures.",
keywords = "Classification, Cryo-electron microscopy, Heterogeneous mixtures, Image processing, Methods development, Multiple structures",
author = "Verbeke, {Eric J.} and Yi Zhou and Horton, {Andrew P.} and Mallam, {Anna L.} and Taylor, {David W.} and Marcotte, {Edward M.}",
note = "Funding Information: We thank S. Musalgaonkar and A. Johnson for providing 40S and 60S yeast ribosomes; A. Dai for expert cryo-EM assistance at the Sauer Structural Biology Laboratory at UT Austin; and members of the Taylor and Marcotte laboratories for helpful discussions. This work was supported in part by Welch Foundation Research Grants F-1938 (to D.W.T.) and F-1515 (to E.M.M.), Army Research Office Grant W911NF-15-1-0120 (to D.W.T.), a Robert J. Kleberg, Jr. and Helen C. Kleberg Foundation Medical Research Award (to D.W.T.), and grants from the National Institutes of Health ( GM122480 , DK110520 , HD085901 ) to E.M.M.. D.W.T is a CPRIT Scholar supported by the Cancer Prevention and Research Institute of Texas (RR160088) and an Army Young Investigator supported by the Army Research Office (W911NF-19-1-0021). Appendix A Funding Information: We thank S. Musalgaonkar and A. Johnson for providing 40S and 60S yeast ribosomes; A. Dai for expert cryo-EM assistance at the Sauer Structural Biology Laboratory at UT Austin; and members of the Taylor and Marcotte laboratories for helpful discussions. This work was supported in part by Welch Foundation Research Grants F-1938 (to D.W.T.) and F-1515 (to E.M.M.), Army Research Office Grant W911NF-15-1-0120 (to D.W.T.), a Robert J. Kleberg, Jr. and Helen C. Kleberg Foundation Medical Research Award (to D.W.T.), and grants from the National Institutes of Health (GM122480, DK110520, HD085901) to E.M.M. D.W.T is a CPRIT Scholar supported by the Cancer Prevention and Research Institute of Texas (RR160088) and an Army Young Investigator supported by the Army Research Office (W911NF-19-1-0021). Publisher Copyright: {\textcopyright} 2019 The Author(s)",
year = "2020",
month = jan,
day = "1",
doi = "10.1016/j.jsb.2019.107416",
language = "English (US)",
volume = "209",
journal = "Journal of Structural Biology",
issn = "1047-8477",
publisher = "Academic Press",
number = "1",
}