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Small stepping motion of processive dynein revealed by load-free high-speed single-particle tracking
Cytoplasmic dynein is a dimeric motor protein which processively moves along microtubule. Its motor domain (head) hydrolyzes ATP and induces conformational changes of linker, stalk, and microtubule binding domain (MTBD) to trigger stepping motion. Here we applied scattering imaging of gold nanoparti...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978368/ https://www.ncbi.nlm.nih.gov/pubmed/31974448 http://dx.doi.org/10.1038/s41598-020-58070-y |
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author | Ando, Jun Shima, Tomohiro Kanazawa, Riko Shimo-Kon, Rieko Nakamura, Akihiko Yamamoto, Mayuko Kon, Takahide Iino, Ryota |
author_facet | Ando, Jun Shima, Tomohiro Kanazawa, Riko Shimo-Kon, Rieko Nakamura, Akihiko Yamamoto, Mayuko Kon, Takahide Iino, Ryota |
author_sort | Ando, Jun |
collection | PubMed |
description | Cytoplasmic dynein is a dimeric motor protein which processively moves along microtubule. Its motor domain (head) hydrolyzes ATP and induces conformational changes of linker, stalk, and microtubule binding domain (MTBD) to trigger stepping motion. Here we applied scattering imaging of gold nanoparticle (AuNP) to visualize load-free stepping motion of processive dynein. We observed artificially-dimerized chimeric dynein, which has the head, linker, and stalk from Dictyostelium discoideum cytoplasmic dynein and the MTBD from human axonemal dynein, whose structure has been well-studied by cryo-electron microscopy. One head of a dimer was labeled with 30 nm AuNP, and stepping motions were observed with 100 μs time resolution and sub-nanometer localization precision at physiologically-relevant 1 mM ATP. We found 8 nm forward and backward steps and 5 nm side steps, consistent with on- and off-axes pitches of binding cleft between αβ-tubulin dimers on the microtubule. Probability of the forward step was 1.8 times higher than that of the backward step, and similar to those of the side steps. One-head bound states were not clearly observed, and the steps were limited by a single rate constant. Our results indicate dynein mainly moves with biased small stepping motion in which only backward steps are slightly suppressed. |
format | Online Article Text |
id | pubmed-6978368 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-69783682020-01-30 Small stepping motion of processive dynein revealed by load-free high-speed single-particle tracking Ando, Jun Shima, Tomohiro Kanazawa, Riko Shimo-Kon, Rieko Nakamura, Akihiko Yamamoto, Mayuko Kon, Takahide Iino, Ryota Sci Rep Article Cytoplasmic dynein is a dimeric motor protein which processively moves along microtubule. Its motor domain (head) hydrolyzes ATP and induces conformational changes of linker, stalk, and microtubule binding domain (MTBD) to trigger stepping motion. Here we applied scattering imaging of gold nanoparticle (AuNP) to visualize load-free stepping motion of processive dynein. We observed artificially-dimerized chimeric dynein, which has the head, linker, and stalk from Dictyostelium discoideum cytoplasmic dynein and the MTBD from human axonemal dynein, whose structure has been well-studied by cryo-electron microscopy. One head of a dimer was labeled with 30 nm AuNP, and stepping motions were observed with 100 μs time resolution and sub-nanometer localization precision at physiologically-relevant 1 mM ATP. We found 8 nm forward and backward steps and 5 nm side steps, consistent with on- and off-axes pitches of binding cleft between αβ-tubulin dimers on the microtubule. Probability of the forward step was 1.8 times higher than that of the backward step, and similar to those of the side steps. One-head bound states were not clearly observed, and the steps were limited by a single rate constant. Our results indicate dynein mainly moves with biased small stepping motion in which only backward steps are slightly suppressed. Nature Publishing Group UK 2020-01-23 /pmc/articles/PMC6978368/ /pubmed/31974448 http://dx.doi.org/10.1038/s41598-020-58070-y Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Ando, Jun Shima, Tomohiro Kanazawa, Riko Shimo-Kon, Rieko Nakamura, Akihiko Yamamoto, Mayuko Kon, Takahide Iino, Ryota Small stepping motion of processive dynein revealed by load-free high-speed single-particle tracking |
title | Small stepping motion of processive dynein revealed by load-free high-speed single-particle tracking |
title_full | Small stepping motion of processive dynein revealed by load-free high-speed single-particle tracking |
title_fullStr | Small stepping motion of processive dynein revealed by load-free high-speed single-particle tracking |
title_full_unstemmed | Small stepping motion of processive dynein revealed by load-free high-speed single-particle tracking |
title_short | Small stepping motion of processive dynein revealed by load-free high-speed single-particle tracking |
title_sort | small stepping motion of processive dynein revealed by load-free high-speed single-particle tracking |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978368/ https://www.ncbi.nlm.nih.gov/pubmed/31974448 http://dx.doi.org/10.1038/s41598-020-58070-y |
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