+
Open data
-
Basic information
Entry | Database: EMDB / ID: EMD-1702 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Kif18A (ATP state) head bound to a microtubule | |||||||||
![]() | 15 protofilament microtubule decorated with the motor domain of Kif18A complexed with AMPPNP | |||||||||
![]() |
| |||||||||
![]() | Molecular motor / kinesin 8 / ruby-helix | |||||||||
Biological species | ![]() ![]() ![]() | |||||||||
Method | helical reconstruction / cryo EM / Resolution: 13.0 Å | |||||||||
![]() | Peters C / Brejc K / Belmont L / Bodey A / Lee Y / Yu M / Ramchandani S / Guo J / Lichtsteiner S / Wood KW ...Peters C / Brejc K / Belmont L / Bodey A / Lee Y / Yu M / Ramchandani S / Guo J / Lichtsteiner S / Wood KW / Sakowicz R / Hartman J / Moores C | |||||||||
![]() | ![]() Title: Insight into the molecular mechanism of the multitasking kinesin-8 motor. Authors: Carsten Peters / Katjuša Brejc / Lisa Belmont / Andrew J Bodey / Yan Lee / Ming Yu / Jun Guo / Roman Sakowicz / James Hartman / Carolyn A Moores / ![]() Abstract: Members of the kinesin-8 motor class have the remarkable ability to both walk towards microtubule plus-ends and depolymerise these ends on arrival, thereby regulating microtubule length. To analyse ...Members of the kinesin-8 motor class have the remarkable ability to both walk towards microtubule plus-ends and depolymerise these ends on arrival, thereby regulating microtubule length. To analyse how kinesin-8 multitasks, we studied the structure and function of the kinesin-8 motor domain. We determined the first crystal structure of a kinesin-8 and used cryo-electron microscopy to calculate the structure of the microtubule-bound motor. Microtubule-bound kinesin-8 reveals a new conformation compared with the crystal structure, including a bent conformation of the α4 relay helix and ordering of functionally important loops. The kinesin-8 motor domain does not depolymerise stabilised microtubules with ATP but does form tubulin rings in the presence of a non-hydrolysable ATP analogue. This shows that, by collaborating, kinesin-8 motor domain molecules can release tubulin from microtubules, and that they have a similar mechanical effect on microtubule ends as kinesin-13, which enables depolymerisation. Our data reveal aspects of the molecular mechanism of kinesin-8 motors that contribute to their unique dual motile and depolymerising functions, which are adapted to control microtubule length. | |||||||||
History |
|
-
Structure visualization
Movie |
![]() |
---|---|
Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
-
Downloads & links
-EMDB archive
Map data | ![]() | 107.3 MB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 10.8 KB 10.8 KB | Display Display | ![]() |
Images | ![]() | 732.8 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
-
Links
EMDB pages | ![]() ![]() |
---|
-
Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | 15 protofilament microtubule decorated with the motor domain of Kif18A complexed with AMPPNP | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.4 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-
Sample components
-Entire : Microtubule complexed with motor domain of Kif18A (ATP state)
Entire | Name: Microtubule complexed with motor domain of Kif18A (ATP state) |
---|---|
Components |
|
-Supramolecule #1000: Microtubule complexed with motor domain of Kif18A (ATP state)
Supramolecule | Name: Microtubule complexed with motor domain of Kif18A (ATP state) type: sample / ID: 1000 / Number unique components: 3 |
---|
-Macromolecule #1: Motor domain of Kif18A
Macromolecule | Name: Motor domain of Kif18A / type: protein_or_peptide / ID: 1 / Name.synonym: Motor domain of Kif18A / Details: Contains the N-terminal 355 aa / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() |
Recombinant expression | Organism: Escherichia coli Rosetta DE3 |
-Macromolecule #2: Beta tubulin
Macromolecule | Name: Beta tubulin / type: protein_or_peptide / ID: 2 / Name.synonym: Beta tubulin / Recombinant expression: No / Database: NCBI |
---|---|
Source (natural) | Organism: ![]() ![]() |
-Macromolecule #3: Alpha tubulin
Macromolecule | Name: Alpha tubulin / type: protein_or_peptide / ID: 3 / Name.synonym: Alpha tubulin / Recombinant expression: No / Database: NCBI |
---|---|
Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
![]() | helical reconstruction |
Aggregation state | filament |
-
Sample preparation
Buffer | pH: 6.8 Details: 80mM PIPES, 150mM NaCl, 7mM MgCl2, 1mM EGTA, 1mM beta-mercaptoethanol |
---|---|
Grid | Details: 400 mesh copper grid |
Vitrification | Cryogen name: ETHANE / Instrument: HOMEMADE PLUNGER / Details: Vitrification instrument: Manual plunger / Timed resolved state: Sample contained 2mM AMPPNP / Method: Blotted for 1 sec before plunging |
-
Electron microscopy
Microscope | FEI TECNAI F20 |
---|---|
Alignment procedure | Legacy - Astigmatism: Objective lense stigmatism was corrected at 150,000 times magnification |
Details | Low dose |
Image recording | Category: FILM / Film or detector model: KODAK SO-163 FILM / Digitization - Scanner: ZEISS SCAI / Digitization - Sampling interval: 7 µm / Number real images: 21 / Average electron dose: 10 e/Å2 / Bits/pixel: 12 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: OTHER / Cs: 2.0 mm / Nominal defocus max: 1.82 µm / Nominal defocus min: 0.74 µm / Nominal magnification: 50000 |
Sample stage | Specimen holder: Cryo-holder / Specimen holder model: GATAN LIQUID NITROGEN |
Experimental equipment | ![]() Model: Tecnai F20 / Image courtesy: FEI Company |
-
Image processing
Final reconstruction | Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 13.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: Ruby-Helix Details: Fourier Bessel Synthesis using data from 24 microtubules (32000 asymmetric units) |
---|---|
CTF correction | Details: Phase flipping, Wiener |