skip to main content

DOE PAGESDOE PAGES

Title: A new protocol to accurately determine microtubule lattice seam location

Microtubules (MTs) are cylindrical polymers of αβ-tubulin that display pseudo-helical symmetry due to the presence of a lattice seam of heterologous lateral contacts. The structural similarity between α- and β-tubulin makes it difficult to computationally distinguish them in the noisy cryo-EM images, unless a marker protein for the tubulin dimer, such as kinesin motor domain, is present. We have developed a new data processing protocol that can accurately determine αβ-tubulin register and seam location for MT segments. Our strategy can handle difficult situations, where the marker protein is relatively small or the decoration of marker protein is sparse. Using this new seam-search protocol, combined with movie processing for data from a direct electron detection camera, we were able to determine the cryo-EM structures of MT at 3.5. Å resolution in different functional states. The successful distinction of α- and β-tubulin allowed us to visualize the nucleotide state at the E-site and the configuration of lateral contacts at the seam.
Authors:
 [1] ;  [1]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States)
Publication Date:
Grant/Contract Number:
AC02-05CH11231
Type:
Accepted Manuscript
Journal Name:
Journal of Structural Biology
Additional Journal Information:
Journal Volume: 192; Journal Issue: 2; Journal ID: ISSN 1047-8477
Publisher:
Elseiver
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
National Institutes of Health (NIH); USDOE
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1378621

Zhang, Rui, and Nogales, Eva. A new protocol to accurately determine microtubule lattice seam location. United States: N. p., Web. doi:10.1016/j.jsb.2015.09.015.
Zhang, Rui, & Nogales, Eva. A new protocol to accurately determine microtubule lattice seam location. United States. doi:10.1016/j.jsb.2015.09.015.
Zhang, Rui, and Nogales, Eva. 2015. "A new protocol to accurately determine microtubule lattice seam location". United States. doi:10.1016/j.jsb.2015.09.015. https://www.osti.gov/servlets/purl/1378621.
@article{osti_1378621,
title = {A new protocol to accurately determine microtubule lattice seam location},
author = {Zhang, Rui and Nogales, Eva},
abstractNote = {Microtubules (MTs) are cylindrical polymers of αβ-tubulin that display pseudo-helical symmetry due to the presence of a lattice seam of heterologous lateral contacts. The structural similarity between α- and β-tubulin makes it difficult to computationally distinguish them in the noisy cryo-EM images, unless a marker protein for the tubulin dimer, such as kinesin motor domain, is present. We have developed a new data processing protocol that can accurately determine αβ-tubulin register and seam location for MT segments. Our strategy can handle difficult situations, where the marker protein is relatively small or the decoration of marker protein is sparse. Using this new seam-search protocol, combined with movie processing for data from a direct electron detection camera, we were able to determine the cryo-EM structures of MT at 3.5. Å resolution in different functional states. The successful distinction of α- and β-tubulin allowed us to visualize the nucleotide state at the E-site and the configuration of lateral contacts at the seam.},
doi = {10.1016/j.jsb.2015.09.015},
journal = {Journal of Structural Biology},
number = 2,
volume = 192,
place = {United States},
year = {2015},
month = {9}
}