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- EMDB-25119: Cryo-EM structure of MAP7 MTBD and microtubule-associated protein... -

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Basic information

Entry
Database: EMDB / ID: EMD-25119
TitleCryo-EM structure of MAP7 MTBD and microtubule-associated protein tau, bound to the microtubule
Map data
Sample
  • Complex: complex of alpha-beta tubulin with MAP7 MTBD and microtubule associated protein tau
Biological speciesSus scrofa (pig)
Methodhelical reconstruction / cryo EM / Resolution: 4.0 Å
AuthorsFerro LS / Fang Q / Eshun-Wilson L / Fernandes J / Jack A / Farrell DP / Golcuk M / Huijben T / Costa K / Gur M ...Ferro LS / Fang Q / Eshun-Wilson L / Fernandes J / Jack A / Farrell DP / Golcuk M / Huijben T / Costa K / Gur M / DiMaio F / Nogales E / Yildiz A
Funding support United States, 7 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM094522 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM123655-03 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM051487 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM127018 United States
National Science Foundation (NSF, United States)MCB-1617028 United States
National Science Foundation (NSF, United States)MCB-1055017 United States
Howard Hughes Medical Institute (HHMI) United States
CitationJournal: Science / Year: 2022
Title: Structural and functional insight into regulation of kinesin-1 by microtubule-associated protein MAP7.
Authors: Luke S Ferro / Qianglin Fang / Lisa Eshun-Wilson / Jonathan Fernandes / Amanda Jack / Daniel P Farrell / Mert Golcuk / Teun Huijben / Katelyn Costa / Mert Gur / Frank DiMaio / Eva Nogales / Ahmet Yildiz /
Abstract: Microtubule (MT)-associated protein 7 (MAP7) is a required cofactor for kinesin-1-driven transport of intracellular cargoes. Using cryo-electron microscopy and single-molecule imaging, we ...Microtubule (MT)-associated protein 7 (MAP7) is a required cofactor for kinesin-1-driven transport of intracellular cargoes. Using cryo-electron microscopy and single-molecule imaging, we investigated how MAP7 binds MTs and facilitates kinesin-1 motility. The MT-binding domain (MTBD) of MAP7 bound MTs as an extended α helix between the protofilament ridge and the site of lateral contact. Unexpectedly, the MTBD partially overlapped with the binding site of kinesin-1 and inhibited its motility. However, by tethering kinesin-1 to the MT, the projection domain of MAP7 prevented dissociation of the motor and facilitated its binding to available neighboring sites. The inhibitory effect of the MTBD dominated as MTs became saturated with MAP7. Our results reveal biphasic regulation of kinesin-1 by MAP7 in the context of their competitive binding to MTs.
History
DepositionOct 7, 2021-
Header (metadata) releaseMay 18, 2022-
Map releaseMay 18, 2022-
UpdateNov 30, 2022-
Current statusNov 30, 2022Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_25119.png

Downloads & links

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Map

FileDownload / File: emd_25119.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.14 Å/pix.
x 512 pix.
= 583.68 Å		1.14 Å/pix.
x 512 pix.
= 583.68 Å		1.14 Å/pix.
x 512 pix.
= 583.68 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.14 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 1.6
Minimum - Maximum-5.478922 - 15.21245
Average (Standard dev.)-0.0754577 (±0.94081175)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-256-256-256
Dimensions512512512
Spacing512512512
CellA=B=C: 583.68 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : complex of alpha-beta tubulin with MAP7 MTBD and microtubule asso...

EntireName: complex of alpha-beta tubulin with MAP7 MTBD and microtubule associated protein tau
Components
  • Complex: complex of alpha-beta tubulin with MAP7 MTBD and microtubule associated protein tau

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Supramolecule #1: complex of alpha-beta tubulin with MAP7 MTBD and microtubule asso...

SupramoleculeName: complex of alpha-beta tubulin with MAP7 MTBD and microtubule associated protein tau
type: complex / ID: 1 / Parent: 0
Source (natural)Organism: Sus scrofa (pig)

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Experimental details

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Structure determination

Methodcryo EM
Processinghelical reconstruction
Aggregation statefilament

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Sample preparation

BufferpH: 7
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TALOS ARCTICA
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 50.0 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

Final reconstructionApplied symmetry - Helical parameters - Δz: 9.483 Å
Applied symmetry - Helical parameters - Δ&Phi: -27.667 °
Applied symmetry - Helical parameters - Axial symmetry: C1 (asymmetric)
Resolution.type: BY AUTHOR / Resolution: 4.0 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 37442
Final angle assignmentType: MAXIMUM LIKELIHOOD

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