+Open data
-Basic information
Entry | Database: PDB / ID: 6ez8 | ||||||||||||
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Title | Human Huntingtin-HAP40 complex structure | ||||||||||||
Components |
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Keywords | PROTEIN BINDING / Huntingtin / HAP40/F8A / Cryo-EM / Huntington's disease | ||||||||||||
Function / homology | Function and homology information vesicle cytoskeletal trafficking / regulation of cAMP-dependent protein kinase activity / : / positive regulation of inositol 1,4,5-trisphosphate-sensitive calcium-release channel activity / microtubule-based transport / vocal learning / regulation of CAMKK-AMPK signaling cascade / negative regulation of proteasomal protein catabolic process / positive regulation of mitophagy / profilin binding ...vesicle cytoskeletal trafficking / regulation of cAMP-dependent protein kinase activity / : / positive regulation of inositol 1,4,5-trisphosphate-sensitive calcium-release channel activity / microtubule-based transport / vocal learning / regulation of CAMKK-AMPK signaling cascade / negative regulation of proteasomal protein catabolic process / positive regulation of mitophagy / profilin binding / vesicle transport along microtubule / positive regulation of cilium assembly / retrograde vesicle-mediated transport, Golgi to endoplasmic reticulum / presynaptic cytosol / positive regulation of aggrephagy / positive regulation of lipophagy / dynein intermediate chain binding / postsynaptic cytosol / beta-tubulin binding / Golgi organization / establishment of mitotic spindle orientation / dynactin binding / Regulation of MECP2 expression and activity / autophagosome / inclusion body / heat shock protein binding / centriole / negative regulation of extrinsic apoptotic signaling pathway / kinase binding / protein destabilization / cytoplasmic vesicle membrane / p53 binding / late endosome / transmembrane transporter binding / early endosome / nuclear body / positive regulation of apoptotic process / axon / dendrite / apoptotic process / perinuclear region of cytoplasm / Golgi apparatus / endoplasmic reticulum / protein-containing complex / nucleoplasm / identical protein binding / nucleus / cytoplasm / cytosol Similarity search - Function | ||||||||||||
Biological species | Homo sapiens (human) | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4 Å | ||||||||||||
Authors | Guo, Q. / Bin, H. / Cheng, J. / Pfeifer, G. / Baumeister, W. / Fernandez-Busnadiego, R. / Kochanek, S. | ||||||||||||
Funding support | Germany, United States, 3items
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Citation | Journal: Nature / Year: 2018 Title: The cryo-electron microscopy structure of huntingtin. Authors: Qiang Guo / Bin Huang / Jingdong Cheng / Manuel Seefelder / Tatjana Engler / Günter Pfeifer / Patrick Oeckl / Markus Otto / Franziska Moser / Melanie Maurer / Alexander Pautsch / Wolfgang ...Authors: Qiang Guo / Bin Huang / Jingdong Cheng / Manuel Seefelder / Tatjana Engler / Günter Pfeifer / Patrick Oeckl / Markus Otto / Franziska Moser / Melanie Maurer / Alexander Pautsch / Wolfgang Baumeister / Rubén Fernández-Busnadiego / Stefan Kochanek / Abstract: Huntingtin (HTT) is a large (348 kDa) protein that is essential for embryonic development and is involved in diverse cellular activities such as vesicular transport, endocytosis, autophagy and the ...Huntingtin (HTT) is a large (348 kDa) protein that is essential for embryonic development and is involved in diverse cellular activities such as vesicular transport, endocytosis, autophagy and the regulation of transcription. Although an integrative understanding of the biological functions of HTT is lacking, the large number of identified HTT interactors suggests that it serves as a protein-protein interaction hub. Furthermore, Huntington's disease is caused by a mutation in the HTT gene, resulting in a pathogenic expansion of a polyglutamine repeat at the amino terminus of HTT. However, only limited structural information regarding HTT is currently available. Here we use cryo-electron microscopy to determine the structure of full-length human HTT in a complex with HTT-associated protein 40 (HAP40; encoded by three F8A genes in humans) to an overall resolution of 4 Å. HTT is largely α-helical and consists of three major domains. The amino- and carboxy-terminal domains contain multiple HEAT (huntingtin, elongation factor 3, protein phosphatase 2A and lipid kinase TOR) repeats arranged in a solenoid fashion. These domains are connected by a smaller bridge domain containing different types of tandem repeats. HAP40 is also largely α-helical and has a tetratricopeptide repeat-like organization. HAP40 binds in a cleft and contacts the three HTT domains by hydrophobic and electrostatic interactions, thereby stabilizing the conformation of HTT. These data rationalize previous biochemical results and pave the way for improved understanding of the diverse cellular functions of HTT. | ||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 6ez8.cif.gz | 467.6 KB | Display | PDBx/mmCIF format |
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PDB format | pdb6ez8.ent.gz | 379.2 KB | Display | PDB format |
PDBx/mmJSON format | 6ez8.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 6ez8_validation.pdf.gz | 860.1 KB | Display | wwPDB validaton report |
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Full document | 6ez8_full_validation.pdf.gz | 878.7 KB | Display | |
Data in XML | 6ez8_validation.xml.gz | 66.3 KB | Display | |
Data in CIF | 6ez8_validation.cif.gz | 102.5 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/ez/6ez8 ftp://data.pdbj.org/pub/pdb/validation_reports/ez/6ez8 | HTTPS FTP |
-Related structure data
Related structure data | 3984MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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-Components
#1: Protein | Mass: 347475.375 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: HTT, HD, IT15 / Production host: Homo sapiens (human) / References: UniProt: P42858 |
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#2: Protein | Mass: 39141.879 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: F8A1, F8A, F8A2, F8A, F8A3, F8A / Production host: Homo sapiens (human) / References: UniProt: P23610 |
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Huntingtin-HAP40 complex / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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Molecular weight | Value: 0.352 MDa / Experimental value: YES |
Source (natural) | Organism: Homo sapiens (human) |
Source (recombinant) | Organism: Homo sapiens (human) / Cell: HEK293 |
Buffer solution | pH: 8 |
Specimen | Conc.: 0.5 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Grid material: GOLD |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE-PROPANE / Humidity: 100 % |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Calibrated defocus min: 1400 nm / Calibrated defocus max: 3000 nm |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Electron dose: 32 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
-Processing
Software | Name: PHENIX / Version: 1.12_2829: / Classification: refinement | ||||||||||||||||||||||||
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EM software | Name: RELION / Version: 2.1 / Category: 3D reconstruction | ||||||||||||||||||||||||
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||
3D reconstruction | Resolution: 4 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 98310 / Symmetry type: POINT | ||||||||||||||||||||||||
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