[English] 日本語
Yorodumi
- EMDB-8169: cryo-EM map of the full-length human NPC1 in complex with the cle... -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: EMDB / ID: EMD-8169
Titlecryo-EM map of the full-length human NPC1 in complex with the cleaved glycoprotein of Ebola virus
Map dataNone
Sample
  • Complex: NPC1 in complex with EBOV-GPcl trimer
    • Protein or peptide: NPC1 in complex with EBOV-GPcl trimer
Function / homology
Function and homology information


cyclodextrin metabolic process / cholesterol storage / membrane raft organization / : / intracellular cholesterol transport / intestinal cholesterol absorption / LDL clearance / cholesterol efflux / programmed cell death / negative regulation of epithelial cell apoptotic process ...cyclodextrin metabolic process / cholesterol storage / membrane raft organization / : / intracellular cholesterol transport / intestinal cholesterol absorption / LDL clearance / cholesterol efflux / programmed cell death / negative regulation of epithelial cell apoptotic process / cholesterol transport / bile acid metabolic process / establishment of protein localization to membrane / adult walking behavior / lysosomal transport / cholesterol binding / negative regulation of macroautophagy / cellular response to steroid hormone stimulus / protein glycosylation / cellular response to low-density lipoprotein particle stimulus / response to cadmium ion / negative regulation of TORC1 signaling / cholesterol metabolic process / neurogenesis / cholesterol homeostasis / liver development / macroautophagy / autophagy / endocytosis / transmembrane signaling receptor activity / virus receptor activity / gene expression / signaling receptor activity / nuclear envelope / late endosome membrane / clathrin-dependent endocytosis of virus by host cell / suppression by virus of host tetherin activity / entry receptor-mediated virion attachment to host cell / lysosome / response to xenobiotic stimulus / symbiont entry into host cell / membrane raft / lysosomal membrane / fusion of virus membrane with host endosome membrane / viral envelope / lipid binding / symbiont-mediated suppression of host type I interferon-mediated signaling pathway / host cell plasma membrane / virion membrane / perinuclear region of cytoplasm / Golgi apparatus / endoplasmic reticulum / extracellular exosome / extracellular region / membrane / plasma membrane
Similarity search - Function
NPC1-like / Niemann-Pick C1, N-terminal / Niemann-Pick C1 N terminus / Filoviruses glycoprotein, extracellular domain / Filoviruses glycoprotein / Filovirus glycoprotein / Sterol-sensing domain of SREBP cleavage-activation / Protein patched/dispatched / Patched family / Sterol-sensing domain (SSD) profile. / Sterol-sensing domain
Similarity search - Domain/homology
NPC intracellular cholesterol transporter 1 / Envelope glycoprotein
Similarity search - Component
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 6.56 Å
AuthorsYan N / Zhou Q / Gong X / Qiang HW / Zhou XH / Wu JP / Gao F / Wan T / Shi Y
Funding support China, 5 items
OrganizationGrant numberCountry
Ministry of Science and Technology of China2015CB9101012014 China
National Natural Science Foundation of China31321062 China
Ministry of Science and Technology of ChinaZX09507003006 China
special project of Ebola virus research from the President Foundation of Chinese Academy of Sciences and Strategic Priority Research Program of the Chinese Academy of SciencesXDB08020100 China
National Natural Science Foundation of China81590761 China
CitationJournal: Cell / Year: 2016
Title: Structural Insights into the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer and Ebola Infection.
Authors: Xin Gong / Hongwu Qian / Xinhui Zhou / Jianping Wu / Tao Wan / Pingping Cao / Weiyun Huang / Xin Zhao / Xudong Wang / Peiyi Wang / Yi Shi / George F Gao / Qiang Zhou / Nieng Yan /
Abstract: Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. ...Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. NPC1 is also the intracellular receptor for Ebola virus (EBOV). Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. A trimeric EBOV-GPcl binds to one NPC1 monomer through the domain C. Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection.
History
DepositionMay 1, 2016-
Header (metadata) releaseJun 8, 2016-
Map releaseJun 8, 2016-
UpdateAug 5, 2020-
Current statusAug 5, 2020Processing site: PDBj / Status: Released

-
Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.02
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by height
  • Surface level: 0.02
  • Imaged by UCSF Chimera
  • Download
  • Surface view with fitted model
  • Atomic models: PDB-5jnx
  • Surface level: 0.02
  • Imaged by UCSF Chimera
  • Download
  • Simplified surface model + fitted atomic model
  • Atomic modelsPDB-5jnx
  • Imaged by Jmol
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

-
Map

FileDownload / File: emd_8169.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationNone
Voxel sizeX=Y=Z: 1.30654 Å
Density
Contour LevelBy AUTHOR: 0.02 / Movie #1: 0.02
Minimum - Maximum-0.041813944 - 0.09011536
Average (Standard dev.)0.00011840048 (±0.0043782326)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 334.47424 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.30653906251.30653906251.3065390625
M x/y/z256256256
origin x/y/z0.0000.0000.000
length x/y/z334.474334.474334.474
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ352352352
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS256256256
D min/max/mean-0.0420.0900.000

-
Supplemental data

-
Sample components

-
Entire : NPC1 in complex with EBOV-GPcl trimer

EntireName: NPC1 in complex with EBOV-GPcl trimer
Components
  • Complex: NPC1 in complex with EBOV-GPcl trimer
    • Protein or peptide: NPC1 in complex with EBOV-GPcl trimer

-
Supramolecule #1: NPC1 in complex with EBOV-GPcl trimer

SupramoleculeName: NPC1 in complex with EBOV-GPcl trimer / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Homo sapiens (human) / Organ: lysosome
Recombinant expressionOrganism: Homo sapiens (human) / Recombinant plasmid: pCAG
Molecular weightTheoretical: 300 KDa

-
Macromolecule #1: NPC1 in complex with EBOV-GPcl trimer

MacromoleculeName: NPC1 in complex with EBOV-GPcl trimer / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MTARGLALGL LLLLLCPAQV FSQSCVWYGE CGIAYGDKRY NCEYSGPPKP LPKDGYDLVQ ELCPGFFFGN VSLCCDVRQL QTLKDNLQL PLQFLSRCPS CFYNLLNLFC ELTCSPRQSQ FLNVTATEDY VDPVTNQTKT NVKELQYYVG QSFANAMYNA C RDVEAPSS ...String:
MTARGLALGL LLLLLCPAQV FSQSCVWYGE CGIAYGDKRY NCEYSGPPKP LPKDGYDLVQ ELCPGFFFGN VSLCCDVRQL QTLKDNLQL PLQFLSRCPS CFYNLLNLFC ELTCSPRQSQ FLNVTATEDY VDPVTNQTKT NVKELQYYVG QSFANAMYNA C RDVEAPSS NDKALGLLCG KDADACNATN WIEYMFNKDN GQAPFTITPV FSDFPVHGME PMNNATKGCD ESVDEVTAPC SC QDCSIVC GPKPQPPPPP APWTILGLDA MYVIMWITYM AFLLVFFGAF FAVWCYRKRY FVSEYTPIDS NIAFSVNASD KGE ASCCDP VSAAFEGCLR RLFTRWGSFC VRNPGCVIFF SLVFITACSS GLVFVRVTTN PVDLWSAPSS QARLEKEYFD QHFG PFFRT EQLIIRAPLT DKHIYQPYPS GADVPFGPPL DIQILHQVLD LQIAIENITA SYDNETVTLQ DICLAPLSPY NTNCT ILSV LNYFQNSHSV LDHKKGDDFF VYADYHTHFL YCVRAPASLN DTSLLHDPCL GTFGGPVFPW LVLGGYDDQN YNNATA LVI TFPVNNYYND TEKLQRAQAW EKEFINFVKN YKNPNLTISF TAERSIEDEL NRESDSDVFT VVISYAIMFL YISLALG HM KSCRRLLVDS KVSLGIAGIL IVLSSVACSL GVFSYIGLPL TLIVIEVIPF LVLAVGVDNI FILVQAYQRD ERLQGETL D QQLGRVLGEV APSMFLSSFS ETVAFFLGAL SVMPAVHTFS LFAGLAVFID FLLQITCFVS LLGLDIKRQE KNRLDIFCC VRGAEDGTSV QASESCLFRF FKNSYSPLLL KDWMRPIVIA IFVGVLSFSI AVLNKVDIGL DQSLSMPDDS YMVDYFKSIS QYLHAGPPV YFVLEEGHDY TSSKGQNMVC GGMGCNNDSL VQQIFNAAQL DNYTRIGFAP SSWIDDYFDW VKPQSSCCRV D NITDQFCN ASVVDPACVR CRPLTPEGKQ RPQGGDFMRF LPMFLSDNPN PKCGKGGHAA YSSAVNILLG HGTRVGATYF MT YHTVLQT SADFIDALKK ARLIASNVTE TMGINGSAYR VFPYSVFYVF YEQYLTIIDD TIFNLGVSLG AIFLVTMVLL GCE LWSAVI MCATIAMVLV NMFGVMWLWG ISLNAVSLVN LVMSCGISVE FCSHITRAFT VSMKGSRVER AEEALAHMGS SVFS GITLT KFGGIVVLAF AKSQIFQIFY FRMYLAMVLL GATHGLIFLP VLLSYIGPSV NKAKSCATEE RYKGTERERL LNF

-
Experimental details

-
Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

-
Sample preparation

Concentration15 mg/mL
BufferpH: 5.5
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 200
VitrificationCryogen name: ETHANE

-
Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsCalibrated defocus max: 2.7 µm / Calibrated defocus min: 1.7 µm / Calibrated magnification: 38270 / Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 2.7 µm / Nominal defocus min: 1.7 µm / Nominal magnification: 22500
Specialist opticsSpherical aberration corrector: no / Chromatic aberration corrector: no / Energy filter - Name: no
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Dimensions - Width: 7676 pixel / Digitization - Dimensions - Height: 7420 pixel / Digitization - Sampling interval: 5.0 µm / Digitization - Frames/image: 1-32 / Number real images: 1379 / Average exposure time: 8.0 sec. / Average electron dose: 50.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

-
Image processing

Particle selectionNumber selected: 703336
CTF correctionSoftware - Name: Gctf (ver. 0.38)
Initial angle assignmentType: PROJECTION MATCHING / Software - Name: RELION (ver. 1.4)
Final angle assignmentType: PROJECTION MATCHING / Software - Name: RELION (ver. 1.4)
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 6.56 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 1.4) / Number images used: 50223

-
Atomic model buiding 1

RefinementProtocol: RIGID BODY FIT
Output model

PDB-5jnx:
The 6.6 A cryo-EM structure of the full-length human NPC1 in complex with the cleaved glycoprotein of Ebola virus

+
About Yorodumi

-
News

-
Feb 9, 2022. New format data for meta-information of EMDB entries

New format data for meta-information of EMDB entries

  • Version 3 of the EMDB header file is now the official format.
  • The previous official version 1.9 will be removed from the archive.

Related info.:EMDB header

External links:wwPDB to switch to version 3 of the EMDB data model

-
Aug 12, 2020. Covid-19 info

Covid-19 info

URL: https://pdbj.org/emnavi/covid19.php

New page: Covid-19 featured information page in EM Navigator.

Related info.:Covid-19 info / Mar 5, 2020. Novel coronavirus structure data

+
Mar 5, 2020. Novel coronavirus structure data

Novel coronavirus structure data

Related info.:Yorodumi Speices / Aug 12, 2020. Covid-19 info

External links:COVID-19 featured content - PDBj / Molecule of the Month (242):Coronavirus Proteases

+
Jan 31, 2019. EMDB accession codes are about to change! (news from PDBe EMDB page)

EMDB accession codes are about to change! (news from PDBe EMDB page)

  • The allocation of 4 digits for EMDB accession codes will soon come to an end. Whilst these codes will remain in use, new EMDB accession codes will include an additional digit and will expand incrementally as the available range of codes is exhausted. The current 4-digit format prefixed with “EMD-” (i.e. EMD-XXXX) will advance to a 5-digit format (i.e. EMD-XXXXX), and so on. It is currently estimated that the 4-digit codes will be depleted around Spring 2019, at which point the 5-digit format will come into force.
  • The EM Navigator/Yorodumi systems omit the EMD- prefix.

Related info.:Q: What is EMD? / ID/Accession-code notation in Yorodumi/EM Navigator

External links:EMDB Accession Codes are Changing Soon! / Contact to PDBj

+
Jul 12, 2017. Major update of PDB

Major update of PDB

  • wwPDB released updated PDB data conforming to the new PDBx/mmCIF dictionary.
  • This is a major update changing the version number from 4 to 5, and with Remediation, in which all the entries are updated.
  • In this update, many items about electron microscopy experimental information are reorganized (e.g. em_software).
  • Now, EM Navigator and Yorodumi are based on the updated data.

External links:wwPDB Remediation / Enriched Model Files Conforming to OneDep Data Standards Now Available in the PDB FTP Archive

-
Yorodumi

Thousand views of thousand structures

  • Yorodumi is a browser for structure data from EMDB, PDB, SASBDB, etc.
  • This page is also the successor to EM Navigator detail page, and also detail information page/front-end page for Omokage search.
  • The word "yorodu" (or yorozu) is an old Japanese word meaning "ten thousand". "mi" (miru) is to see.

Related info.:EMDB / PDB / SASBDB / Comparison of 3 databanks / Yorodumi Search / Aug 31, 2016. New EM Navigator & Yorodumi / Yorodumi Papers / Jmol/JSmol / Function and homology information / Changes in new EM Navigator and Yorodumi

Read more