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- EMDB-10754: Clathrin with bound beta2 appendage of AP2 -

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Basic information

Entry
Database: EMDB / ID: EMD-10754
TitleClathrin with bound beta2 appendage of AP2
Map dataSharpened map of hexagon bounding clathrin leg enriched in beta-appendage
Sample
  • Complex: Clathrin/AP2 coat assembled on membrane.
    • Complex: Clathrin
      • Protein or peptide: Clathrin heavy chain
      • Protein or peptide: Clathrin heavy chain
      • Protein or peptide: Clathrin light chain
    • Complex: beta2 appendage of AP2 adaptor
      • Protein or peptide: AP-2 complex subunit beta
Keywordsclathrin / clathrin adaptor / ap2 / clathrin assembly / ENDOCYTOSIS
Function / homology
Function and homology information


Lysosome Vesicle Biogenesis / WNT5A-dependent internalization of FZD4 / WNT5A-dependent internalization of FZD2, FZD5 and ROR2 / Cargo recognition for clathrin-mediated endocytosis / Gap junction degradation / Formation of annular gap junctions / Clathrin-mediated endocytosis / clathrin coat of trans-Golgi network vesicle / clathrin light chain binding / clathrin complex ...Lysosome Vesicle Biogenesis / WNT5A-dependent internalization of FZD4 / WNT5A-dependent internalization of FZD2, FZD5 and ROR2 / Cargo recognition for clathrin-mediated endocytosis / Gap junction degradation / Formation of annular gap junctions / Clathrin-mediated endocytosis / clathrin coat of trans-Golgi network vesicle / clathrin light chain binding / clathrin complex / Nef Mediated CD8 Down-regulation / clathrin adaptor complex / WNT5A-dependent internalization of FZD2, FZD5 and ROR2 / Trafficking of GluR2-containing AMPA receptors / WNT5A-dependent internalization of FZD4 / clathrin coat of coated pit / AP-2 adaptor complex / postsynaptic neurotransmitter receptor internalization / LDL clearance / clathrin coat assembly / Retrograde neurotrophin signalling / clathrin-coated endocytic vesicle / clathrin-dependent endocytosis / coronary vasculature development / endolysosome membrane / signal sequence binding / Nef Mediated CD4 Down-regulation / aorta development / clathrin-coated vesicle / ventricular septum development / ciliary membrane / clathrin binding / Recycling pathway of L1 / synaptic vesicle endocytosis / EPH-ephrin mediated repulsion of cells / vesicle-mediated transport / MHC class II antigen presentation / VLDLR internalisation and degradation / receptor-mediated endocytosis / kidney development / intracellular protein transport / clathrin-coated endocytic vesicle membrane / trans-Golgi network / cytoplasmic side of plasma membrane / spindle / disordered domain specific binding / endocytic vesicle membrane / Cargo recognition for clathrin-mediated endocytosis / Clathrin-mediated endocytosis / presynapse / mitotic cell cycle / postsynapse / Potential therapeutics for SARS / glutamatergic synapse / nucleolus / structural molecule activity / extracellular exosome / nucleoplasm / membrane / plasma membrane / cytosol
Similarity search - Function
Clathrin light chain / Clathrin light chain signature 1. / Clathrin light chain signature 2. / Clathrin light chain / Clathrin, heavy chain, linker, core motif / Clathrin heavy chain, N-terminal / Clathrin, heavy chain / Clathrin, heavy chain, propeller repeat / Clathrin propeller repeat / Clathrin, heavy-chain linker ...Clathrin light chain / Clathrin light chain signature 1. / Clathrin light chain signature 2. / Clathrin light chain / Clathrin, heavy chain, linker, core motif / Clathrin heavy chain, N-terminal / Clathrin, heavy chain / Clathrin, heavy chain, propeller repeat / Clathrin propeller repeat / Clathrin, heavy-chain linker / Clathrin-H-link / Region in Clathrin and VPS / Clathrin heavy chain repeat homology / Clathrin, heavy chain/VPS, 7-fold repeat / Clathrin heavy-chain (CHCR) repeat profile. / Clathrin adaptor, beta-adaptin, appendage, Ig-like subdomain / Beta-adaptin appendage, C-terminal subdomain / AP-1/2/4 complex subunit beta / Ribosome biogenesis protein Nop16 / Beta2-adaptin appendage, C-terminal sub-domain / Ribosome biogenesis protein Nop16 / Beta2-adaptin appendage, C-terminal sub-domain / AP complex subunit beta / Coatomer/calthrin adaptor appendage, C-terminal subdomain / Clathrin adaptor, alpha/beta/gamma-adaptin, appendage, Ig-like subdomain / Adaptin C-terminal domain / Adaptin C-terminal domain / Clathrin/coatomer adaptor, adaptin-like, N-terminal / Adaptin N terminal region / Clathrin adaptor, appendage, Ig-like subdomain superfamily / TBP domain superfamily / Armadillo/beta-catenin-like repeats / Armadillo / Armadillo-like helical / Tetratricopeptide-like helical domain superfamily / Armadillo-type fold
Similarity search - Domain/homology
Clathrin light chain / Clathrin heavy chain / Nucleolar protein 16 / Clathrin heavy chain / AP-2 complex subunit beta
Similarity search - Component
Biological speciesSus scrofa (pig) / Homo sapiens (human)
Methodsubtomogram averaging / cryo EM / Resolution: 9.2 Å
AuthorsKovtun O / Kane Dickson V
Funding support United Kingdom, 2 items
OrganizationGrant numberCountry
Wellcome Trust207455/Z/17/Z United Kingdom
Medical Research Council (MRC, United Kingdom)MC_UP_1201/16 United Kingdom
CitationJournal: Sci Adv / Year: 2020
Title: Architecture of the AP2/clathrin coat on the membranes of clathrin-coated vesicles.
Authors: Oleksiy Kovtun / Veronica Kane Dickson / Bernard T Kelly / David J Owen / John A G Briggs /
Abstract: Clathrin-mediated endocytosis (CME) is crucial for modulating the protein composition of a cell's plasma membrane. Clathrin forms a cage-like, polyhedral outer scaffold around a vesicle, to which ...Clathrin-mediated endocytosis (CME) is crucial for modulating the protein composition of a cell's plasma membrane. Clathrin forms a cage-like, polyhedral outer scaffold around a vesicle, to which cargo-selecting clathrin adaptors are attached. Adaptor protein complex (AP2) is the key adaptor in CME. Crystallography has shown AP2 to adopt a range of conformations. Here, we used cryo-electron microscopy, tomography, and subtomogram averaging to determine structures, interactions, and arrangements of clathrin and AP2 at the key steps of coat assembly, from AP2 in solution to membrane-assembled clathrin-coated vesicles (CCVs). AP2 binds cargo and PtdIns(4,5) (phosphatidylinositol 4,5-bisphosphate)-containing membranes via multiple interfaces, undergoing conformational rearrangement from its cytosolic state. The binding mode of AP2 β2 appendage into the clathrin lattice in CCVs and buds implies how the adaptor structurally modulates coat curvature and coat disassembly.
History
DepositionMar 12, 2020-
Header (metadata) releaseJul 29, 2020-
Map releaseJul 29, 2020-
UpdateMay 22, 2024-
Current statusMay 22, 2024Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.07
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  • Surface view with fitted model
  • Atomic models: PDB-6yai
  • Surface level: 0.07
  • Imaged by UCSF Chimera
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  • Simplified surface model + fitted atomic model
  • Atomic modelsPDB-6yai
  • Imaged by Jmol
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_10754.png

Downloads & links

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Map

FileDownload / File: emd_10754.map.gz / Format: CCP4 / Size: 15.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSharpened map of hexagon bounding clathrin leg enriched in beta-appendage
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.78 Å/pix.
x 160 pix.
= 284.8 Å		1.78 Å/pix.
x 160 pix.
= 284.8 Å		1.78 Å/pix.
x 160 pix.
= 284.8 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.78 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.07 / Movie #1: 0.07
Minimum - Maximum-0.44512126 - 0.6409551
Average (Standard dev.)-0.00006332948 (±0.055856194)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions160160160
Spacing160160160
CellA=B=C: 284.8 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.781.781.78
M x/y/z160160160
origin x/y/z0.0000.0000.000
length x/y/z284.800284.800284.800
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS160160160
D min/max/mean-0.4450.641-0.000

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Supplemental data

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Mask #1

Fileemd_10754_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Unsharpened half-map of hexagon bounding clathrin leg enriched...

Fileemd_10754_half_map_1.map
AnnotationUnsharpened half-map of hexagon bounding clathrin leg enriched in beta2-appendage
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Unsharpened half-map of hexagon bounding clathrin leg enriched...

Fileemd_10754_half_map_2.map
AnnotationUnsharpened half-map of hexagon bounding clathrin leg enriched in beta-appendage
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Clathrin/AP2 coat assembled on membrane.

EntireName: Clathrin/AP2 coat assembled on membrane.
Components
  • Complex: Clathrin/AP2 coat assembled on membrane.
    • Complex: Clathrin
      • Protein or peptide: Clathrin heavy chain
      • Protein or peptide: Clathrin heavy chain
      • Protein or peptide: Clathrin light chain
    • Complex: beta2 appendage of AP2 adaptor
      • Protein or peptide: AP-2 complex subunit beta

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Supramolecule #1: Clathrin/AP2 coat assembled on membrane.

SupramoleculeName: Clathrin/AP2 coat assembled on membrane. / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Details: Clathrin/AP coat was formed on the membrane containing cargo signal peptides. The AP2 adaptor lacked hinge and appendage regions in its alpha subunit.

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Supramolecule #2: Clathrin

SupramoleculeName: Clathrin / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1, #3-#4
Details: Clathrin in the clathrin/AP2 coat formed on the membrane
Source (natural)Organism: Sus scrofa (pig) / Organ: brain

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Supramolecule #3: beta2 appendage of AP2 adaptor

SupramoleculeName: beta2 appendage of AP2 adaptor / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2
Details: beta2 appendage of the AP2 bound to clathrin to clathrin/AP2 coat formed on the membrane
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: Clathrin heavy chain

MacromoleculeName: Clathrin heavy chain / type: protein_or_peptide / ID: 1 / Number of copies: 8 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
Molecular weightTheoretical: 187.145125 KDa
SequenceString: MAQILPIRFQ EHLQLQNLGI NPANIGFSTL TMESDKFICI REKVGEQAQV VIIDMNDPSN PIRRPISADS AIMNPASKVI ALKAGKTLQ IFNIEMKSKM KAHTMTDDVT FWKWISLNTV ALVTDNAVYH WSMEGESQPV KMFDRHSSLA GCQIINYRTD A KQKWLLLT ...String:
MAQILPIRFQ EHLQLQNLGI NPANIGFSTL TMESDKFICI REKVGEQAQV VIIDMNDPSN PIRRPISADS AIMNPASKVI ALKAGKTLQ IFNIEMKSKM KAHTMTDDVT FWKWISLNTV ALVTDNAVYH WSMEGESQPV KMFDRHSSLA GCQIINYRTD A KQKWLLLT GISAQQNRVV GAMQLYSVDR KVSQPIEGHA ASFAQFKMEG NAEESTLFCF AVRGQAGGKL HIIEVGTPPT GN QPFPKKA VDVFFPPEAQ NDFPVAMQIS EKHDVVFLIT KYGYIHLYDL ETGTCIYMNR ISGETIFVTA PHEATAGIIG VNR KGQVLS VCVEEENIIP YITNVLQNPD LALRMAVRNN LAGAEELFAR KFNALFAQGN YSEAAKVAAN APKGILRTPD TIRR FQSVP AQPGQTSPLL QYFGILLDQG QLNKYESLEL CRPVLQQGRK QLLEKWLKED KLECSEELGD LVKSVDPTLA LSVYL RANV PNKVIQCFAE TGQVQKIVLY AKKVGYTPDW IFLLRNVMRI SPDQGQQFAQ MLVQDEEPLA DITQIVDVFM EYNLIQ QCT AFLLDALKNN RPSEGPLQTR LLEMNLMHAP QVADAILGNQ MFTHYDRAHI AQLCEKAGLL QRALEHFTDL YDIKRAV VH THLLNPEWLV NYFGSLSVED SLECLRAMLS ANIRQNLQIC VQVASKYHEQ LSTQSLIELF ESFKSFEGLF YFLGSIVN F SQDPDVHFKY IQAACKTGQI KEVERICRES NCYDPERVKN FLKEAKLTDQ LPLIIVCDRF DFVHDLVLYL YRNNLQKYI EIYVQKVNPS RLPVVIGGLL DVDCSEDVIK NLILVVRGQF STDELVAEVE KRNRLKLLLP WLEARIHEGC EEPATHNALA KIYIDSNNN PERFLRENPY YDSRVVGKYC EKRDPHLACV AYERGQCDLE LINVCNENSL FKSLSRYLVR RKDPELWGSV L LESNPYRR PLIDQVVQTA LSETQDPEEV SVTVKAFMTA DLPNELIELL EKIVLDNSVF SEHRNLQNLL ILTAIKADRT RV MEYINRL DNYDAPDIAN IAISNELFEE AFAIFRKFDV NTSAVQVLIE HIGNLDRAYE FAERCNEPAV WSQLAKAQLQ KGM VKEAID SYIKADDPSS YMEVVQAANT SGNWEELVKY LQMARKKARE SYVETELIFA LAKTNRLAEL EEFINGPNNA HIQQ VGDRC YDEKMYDAAK LLYNNVSNFG RLASTLVHLG EYQAAVDGAR KANSTRTWKE VCFACVDGKE FRLAQMCGLH IVVHA DELE ELINYYQDRG YFEELITMLE AALGLERAHM GMFTELAILY SKFKPQKMRE HLELFWSRVN IPKVLRAAEQ AHLWAE LVF LYDKYEEYDN AIITMMNHPT DAWKEGQFKD IITKVANVEL YYRAIQFYLE FKPLLLNDLL MVLSPRLDHT RAVNYFS KV KQLPLVKPYL RSVQNHNNKS VNESLNNLFI TEEDYQALRT SIDAYDNFDN ISLAQRLEKH ELIEFRRIAA YLFKGNNR W KQSVELCKKD SLYKDAMQYA SESKDTELAE ELLQWFLQEE KRECFGACLF TCYDLLRPDV VLETAWRHNI MDFAMPYFI QVMKEYLTKV DKLDASESLR KEEEQATETQ

UniProtKB: Clathrin heavy chain

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Macromolecule #2: AP-2 complex subunit beta

MacromoleculeName: AP-2 complex subunit beta / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 26.429457 KDa
Recombinant expressionOrganism: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria)
SequenceString: GGYVAPKAVW LPAVKAKGLE ISGTFTHRQG HIYMEMNFTN KALQHMTDFA IQFNKNSFGV IPSTPLAIHT PLMPNQSIDV SLPLNTLGP VMKMEPLNNL QVAVKNNIDV FYFSCLIPLN VLFVEDGKME RQVFLATWKD IPNENELQFQ IKECHLNADT V SSKLQNNN ...String:
GGYVAPKAVW LPAVKAKGLE ISGTFTHRQG HIYMEMNFTN KALQHMTDFA IQFNKNSFGV IPSTPLAIHT PLMPNQSIDV SLPLNTLGP VMKMEPLNNL QVAVKNNIDV FYFSCLIPLN VLFVEDGKME RQVFLATWKD IPNENELQFQ IKECHLNADT V SSKLQNNN VYTIAKRNVE GQDMLYQSLK LTNGIWILAE LRIQPGNPNY TLSLKCRAPE VSQYIYQVYD SILKN

UniProtKB: AP-2 complex subunit beta

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Macromolecule #3: Clathrin heavy chain

MacromoleculeName: Clathrin heavy chain / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
Molecular weightTheoretical: 187.117094 KDa
SequenceString: MAQILPIRFQ EHLQLQNLGI NPANIGFSTL TMESDKFICI REKVGEQAQV VIIDMNDPSN PIRRPISADS AIMNPASKVI ALKAGKTLQ IFNIEMKSKM KAHTMTDDVT FWKWISLNTV ALVTDNAVYH WSMEGESQPV KMFDRHSSLA GCQIINYRTD A KQKWLLLT ...String:
MAQILPIRFQ EHLQLQNLGI NPANIGFSTL TMESDKFICI REKVGEQAQV VIIDMNDPSN PIRRPISADS AIMNPASKVI ALKAGKTLQ IFNIEMKSKM KAHTMTDDVT FWKWISLNTV ALVTDNAVYH WSMEGESQPV KMFDRHSSLA GCQIINYRTD A KQKWLLLT GISAQQNRVV GAMQLYSVDR KVSQPIEGHA ASFAQFKMEG NAEESTLFCF AVRGQAGGKL HIIEVGTPPT GN QPFPKKA VDVFFPPEAQ NDFPVAMQIS EKHDVVFLIT KYGYIHLYDL ETGTCIYMNR ISGETIFVTA PHEATAGIIG VNR KGQVLS VCVEEENIIP YITNVLQNPD LALRMAVRNN LAGAEELFAR KFNALFAQGN YSEAAKVAAN APKGILRTPD TIRR FQSVP AQPGQTSPLL QYFGILLDQG QLNKYESLEL CRPVLQQGRK QLLEKWLKED KLECSEELGD LVKSVDPTLA LSVYL RANV PNKVIQCFAE TGQVQKIVLY AKKVGYTPDW IFLLRNVMRI SPDQGQQFAQ MLVQDEEPLA DITQIVDVFM EYNLIQ QCT AFLLDALKNN RPSEGPLQTR LLEMNLMHAP QVADAILGNQ MFTHYDRAHI AQLCEKAGLL QRALEHFTDL YDIKRAV VH THLLNPEWLV NYFGSLSVED SLECLRAMLS ANIRQNLQIC VQVASKYHEQ LSTQSLIELF ESFKSFEGLF YFLGSIVN F SQDPDVHFKY IQAACKTGQI KEVERICRES NCYDPERVKN FLKEAKLTDQ LPLIIVCDRF DFVHDLVLYL YRNNLQKYI EIYVQKVNPS RLPVVIGGLL DVDCSEDVIK NLILVVRGQF STDELVAEVE KRNRLKLLLP WLEARIHEGC TEPATHNALA KIYIDSNNN PERFLRENPY YDSRVVGKYC EKRDPHLACV AYERGQCDLE LINVCNENSL FKSLSRYLVR RKDPELWGSV L LESNPYRR PLIDQVVQTA LSETQDPEEV SVTVKAFMTA DLPNELIELL EKIVLDNSVF SEHRNLQNLL ILTAIKADRT RV MEYINRL DNYDAPDIAN IAISNELFEE AFAIFRKFDV NTSAVQVLIE HIGNLDRAYE FAERCNEPAV WSQLAKAQLQ KGM VKEAID SYIKADDPSS YMEVVQAANT SGNWEELVKY LQMARKKARE SYVETELIFA LAKTNRLAEL EEFINGPNNA HIQQ VGDRC YDEKMYDAAK LLYNNVSNFG RLASTLVHLG EYQAAVDGAR KANSTRTWKE VCFACVDGKE FRLAQMCGLH IVVHA DELE ELINYYQDRG YFEELITMLE AALGLERAHM GMFTELAILY SKFKPQKMRE HLELFWSRVN IPKVLRAAEQ AHLWAE LVF LYDKYEEYDN AIITMMNHPT DAWKEGQFKD IITKVANVEL YYRAIQFYLE FKPLLLNDLL MVLSPRLDHT RAVNYFS KV KQLPLVKPYL RSVQNHNNKS VNESLNNLFI TEEDYQALRT SIDAYDNFDN ISLAQRLEKH ELIEFRRIAA YLFKGNNR W KQSVELCKKD SLYKDAMQYA SESKDTELAE ELLQWFLQEE KRECFGACLF TCYDLLRPDV VLETAWRHNI MDFAMPYFI QVMKEYLTKV DKLDASESLR KEEEQATETQ

UniProtKB: Clathrin heavy chain

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Macromolecule #4: Clathrin light chain

MacromoleculeName: Clathrin light chain / type: protein_or_peptide / ID: 4 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
Molecular weightTheoretical: 25.2185 KDa
SequenceString: MADDFGFFSS SESGAPEVAE EDPAAAFLAQ QESEIAGIEN DEGFGAPAGS QAALAQPGPA SGAGPEDMGT TVNGDVFQDA NGPADGYAA IAQADRLTQE PESIRKWREE QRKRLQELDA ASKVTEQEWR EKAKKDLEEW NQRQSEQVEK NKINNRIADK A FYQQPDAD ...String:
MADDFGFFSS SESGAPEVAE EDPAAAFLAQ QESEIAGIEN DEGFGAPAGS QAALAQPGPA SGAGPEDMGT TVNGDVFQDA NGPADGYAA IAQADRLTQE PESIRKWREE QRKRLQELDA ASKVTEQEWR EKAKKDLEEW NQRQSEQVEK NKINNRIADK A FYQQPDAD IIGYVASEEA FVKESKEETP GTEWEKVAQL CDFNPKSSKQ CKDVSRLRSV LMSLKQTPLS R

UniProtKB: Nucleolar protein 16

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation state3D array

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Sample preparation

Concentration0.7 mg/mL
BufferpH: 7.2
Component:
ConcentrationName
10.0 mMHepes KOH
120.0 mMPotassium Acetate
2.0 mMMagnesium chloride
5.0 mM2-Mercaptoethanol
GridModel: C-flat-2/2 / Support film - Material: CARBON / Support film - topology: HOLEY
VitrificationCryogen name: ETHANE / Chamber humidity: 98 % / Chamber temperature: 291 K / Instrument: LEICA EM GP
Details: The sample was supplemented with 10 nm nanogold fiducials, and 3 ul of the mixture was backside blotted for 3 seconds..
DetailsThe sample (in vitro budding reaction) contained AP2, clathrin and 400 nm extruded liposomes

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Specialist opticsEnergy filter - Name: GIF Quantum LS / Energy filter - Slit width: 20 eV
Image recordingFilm or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3838 pixel / Digitization - Dimensions - Height: 3710 pixel / Digitization - Frames/image: 1-10 / Number grids imaged: 2 / Average exposure time: 0.2 sec. / Average electron dose: 3.2 e/Å2
Details: The images were collected in movie mode at 10 frames per second
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Calibrated defocus max: 6.5 µm / Calibrated defocus min: 1.5 µm / Calibrated magnification: 81000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 6.5 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 81000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

DetailsThe images were low pass filtered according to the cumulative radiation dose.
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 9.2 Å / Resolution method: FSC 0.143 CUT-OFF / Software: (Name: TOM, AV3) / Number subtomograms used: 12076
ExtractionNumber tomograms: 58121 / Number images used: 198871 / Reference model: reference-free / Method: geometrically defined initial positions / Software - Name: TOM
Details: To define initial positions of subtomograms, centres and radii of coated vesicles, buds were manually marked in bin4 tomograms. These measurements were used to define spheres. Subtomogram ...Details: To define initial positions of subtomograms, centres and radii of coated vesicles, buds were manually marked in bin4 tomograms. These measurements were used to define spheres. Subtomogram positions were then defined on the surface of these spheres with uniform sampling. The orientations were calculated to be normal to the surfaces of the spheres with random in-plane rotation.
Final 3D classificationNumber classes: 10 / Avg.num./class: 1200 / Software: (Name: PEET, Dynamo)
Details: Principal component analysis (PCA) to sort subtomograms by beta-appendage occupancy. The PCA was performed on wedge-masked difference maps with calculations implemented in MATLAB using code ...Details: Principal component analysis (PCA) to sort subtomograms by beta-appendage occupancy. The PCA was performed on wedge-masked difference maps with calculations implemented in MATLAB using code adapted from PEET and Dynamo packages.
Final angle assignmentType: OTHER / Software: (Name: TOM, AV3)
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial model
PDB IDChain

1xi4

source_name: PDB, initial_model_type: experimental model

6sct

source_name: PDB, initial_model_type: experimental model

1e42

chain_id: A, source_name: PDB, initial_model_type: experimental model
RefinementSpace: REAL / Protocol: FLEXIBLE FIT / Target criteria: correlation coefficient
Output model

PDB-6yai:
Clathrin with bound beta2 appendage of AP2

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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

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