Computational separation of conformational heterogeneity using cryo-electron tomography and 3D sub-volume averaging.

ABSTRACT: We have previously used cryo-electron tomography combined with sub-volume averaging and classification to obtain 3D structures of macromolecular assemblies in cases where a single dominant species was present, and applied these methods to the analysis of a variety of trimeric HIV-1 and SIV envelope glycoproteins (Env). Here, we extend these studies by demonstrating automated, iterative, missing wedge-corrected 3D image alignment and classification methods to distinguish multiple conformations that are present simultaneously. We present a method for measuring the spatial distribution of the vector elements representing distinct conformational states of Env. We identify data processing strategies that allow clear separation of the previously characterized closed and open conformations, as we

SUBMITTER: Frank GA 

PROVIDER: S-EPMC3350607 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature