Detail of Fig4EG_MTelongation_2
Project
Title
Time-series images of x-ray fiber diffraction measuring axial pitch of GDP-MT before and after addition of K11C or AMPPMP
Description
Time-series images of x-ray fiber diffraction measuring axial pitch of GDP-MT (GDP-microtubules) before and after addition of K11C or AMPPMP. K11C is a mutant KIF5C, the L11-alpha4 junction was replaced with the corresponding sequence of KIF1A. A synchrotron radiation x-ray beam at BL45XU beamline of SPring-8 (Japan Synchrotron Radiation Research Institute) irradiated the oriented microtubules. K11C was added after time00001, and AMPPMP was applied after time00011.
Release, Updated
2024-12-14
License
CC BY
Kind
Image data
File Formats
tif
Data size
198.5 MB
Organism
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Strain(s)
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Cell Line
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Datatype
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Molecular Function (MF)
Biological Process (BP)
microtubule bundle maintenance
Cellular Component (CC)
Biological Imaging Method
X scale
0.017 nanometer^{-1}/pixel
Y scale
0.017 nanometer^{-1}/pixel
Z scale
-
T scale
15 seconds per time interval
Image Acquisition
Experiment type
-
Microscope type
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Acquisition mode
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Contrast method
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Microscope model
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Detector model
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Objective model
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Filter set
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Summary of Methods
Shima T, Morikawa M, Kaneshiro J, Kambara T, Kamimura S, Yagi T, Iwamoto H, Uemura S, Shigematsu H, Shirouzu M, Ichimura T, Watanabe TM, Nitta R, Okada Y, Hirokawa N. Kinesin-binding-triggered conformation switching of microtubules contributes to polarized transport. J Cell Biol. 2018 Dec 3;217(12):4164-4183.
Related paper(s)
Tomohiro Shima, Manatsu Morikawa, Junichi Kaneshiro, Taketoshi Kambara, Shinji Kamimura, Toshiki Yagi, Hiroyuki Iwamoto, Sotaro Uemura, Hideki Shigematsu, Mikako Shirouzu, Taro Ichimura, Tomonobu M Watanabe, Ryo Nitta, Yasushi Okada, Nobutaka Hirokawa (2018) Kinesin-binding-triggered conformation switching of microtubules contributes to polarized transport., The Journal of cell biology, Volume 217, Number 12, pp. 4164-4183
Published in 2018 Dec 3 (Electronic publication in Oct. 8, 2018, midnight )
- doi: 10.1083/jcb.201711178
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pmid: 30297389
(Abstract) Kinesin-1, the founding member of the kinesin superfamily of proteins, is known to use only a subset of microtubules for transport in living cells. This biased use of microtubules is proposed as the guidance cue for polarized transport in neurons, but the underlying mechanisms are still poorly understood. Here, we report that kinesin-1 binding changes the microtubule lattice and promotes further kinesin-1 binding. This high-affinity state requires the binding of kinesin-1 in the nucleotide-free state. Microtubules return to the initial low-affinity state by washing out the binding kinesin-1 or by the binding of non-hydrolyzable ATP analogue AMPPNP to kinesin-1. X-ray fiber diffraction, fluorescence speckle microscopy, and second-harmonic generation microscopy, as well as cryo-EM, collectively demonstrated that the binding of nucleotide-free kinesin-1 to GDP microtubules changes the conformation of the GDP microtubule to a conformation resembling the GTP microtubule.(MeSH Terms)
- Adenylyl Imidodiphosphate/chemistry
- Adenylyl Imidodiphosphate/pharmacology
- Animals
- Biological Transport, Active
- Chlorocebus aethiops
- Dogs
- Guanosine Diphosphate/chemistry
- Guanosine Diphosphate/metabolism
- Guanosine Triphosphate/chemistry
- Guanosine Triphosphate/metabolism
- HeLa Cells
- Humans
- Kinesins/chemistry[major]
- Kinesins/metabolism[major]
- Madin Darby Canine Kidney Cells
- Microtubules/chemistry[major]
- Microtubules/metabolism[major]
- Vero Cells
Contact
Yasushi Okada, Nobutaka Hirokawa
, RIKEN BDR, The University of Tokyo
, Laboratory for Cell Polarity Regulation, Department of Cell Biology and Anatomy, Graduate School of Medicine
, Laboratory for Cell Polarity Regulation, Department of Cell Biology and Anatomy, Graduate School of Medicine
Contributors
Tomohiro Shima, Sotaro Uemura, Yasushi Okada
OMERO Project
Source