A new protocol to accurately determine microtubule lattice seam location
Abstract
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:
-
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States)
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- National Institutes of Health (NIH); USDOE
- OSTI Identifier:
- 1378621
- Grant/Contract Number:
- AC02-05CH11231
- Resource 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
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Zhang, Rui, and Nogales, Eva. A new protocol to accurately determine microtubule lattice seam location. United States: N. p., 2015.
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. https://doi.org/10.1016/j.jsb.2015.09.015
Zhang, Rui, and Nogales, Eva. Mon .
"A new protocol to accurately determine microtubule lattice seam location". United States. https://doi.org/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 = {Mon Sep 28 00:00:00 EDT 2015},
month = {Mon Sep 28 00:00:00 EDT 2015}
}
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