Rxivist logo

Template-free detection and classification of heterogeneous membrane-bound complexes in cryo-electron tomograms

By Antonio Martínez-Sánchez, Zdravko Kochovski, Ulrike Laugks, Johannes Meyer zum Alten Borgloh, Saikat Chakraborty, Stefan Pfeffer, Wolfgang Baumeister, Vladan Lucic

Posted 10 Sep 2018
bioRxiv DOI: 10.1101/413484 (published DOI: 10.1038/s41592-019-0675-5)

With faithful sample preservation and direct imaging of fully hydrated biological material, cryo-electron tomography (cryo-ET) provides an accurate representation of molecular architecture of cells. However, detection and precise localization of macromolecular complexes within cellular environments is aggravated by the presence of many molecular species and molecular crowding. We developed a template-free image processing procedure for accurate tracing of complex networks of densities in cryo-electron tomograms, a comprehensive and automated detection of heterogeneous membrane-bound complexes and an unsupervised classification. Applying this procedure to tomograms of intact cells and isolated endoplasmic reticulum (ER), we detected and classified small protein complexes like the ER protein translocons, which were not detected by other methods before. This classification provided sufficiently homogeneous particle sets and initial references to allow subsequent de novo subtomogram averaging. Therefore the procedure presented allows a comprehensive detection and a structural analysis of complexes in their native state. In addition, we present structural evidence that different ribosome-free translocon species are present at the ER membrane, determine their 3D structure, and show that they have different localization patterns forming nanodomains.

Download data

  • Downloaded 1,057 times
  • Download rankings, all-time:
    • Site-wide: 21,929
    • In biophysics: 589
  • Year to date:
    • Site-wide: 52,097
  • Since beginning of last month:
    • Site-wide: 50,443

Altmetric data


Downloads over time

Distribution of downloads per paper, site-wide


PanLingua

News