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- EMDB-22785: Cryo-EM structure of the Sec complex from S. cerevisiae, wild-typ... -
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Basic information
Entry | Database: EMDB / ID: EMD-22785 | |||||||||
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Title | Cryo-EM structure of the Sec complex from S. cerevisiae, wild-type, consensus map | |||||||||
![]() | Unsharpened, lowpass-filtered map | |||||||||
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Function / homology | ![]() misfolded protein transport / Sec62/Sec63 complex / translocon complex / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / cytosol to endoplasmic reticulum transport / rough endoplasmic reticulum membrane / Ssh1 translocon complex / Sec61 translocon complex / protein-transporting ATPase activity / post-translational protein targeting to endoplasmic reticulum membrane ...misfolded protein transport / Sec62/Sec63 complex / translocon complex / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / cytosol to endoplasmic reticulum transport / rough endoplasmic reticulum membrane / Ssh1 translocon complex / Sec61 translocon complex / protein-transporting ATPase activity / post-translational protein targeting to endoplasmic reticulum membrane / filamentous growth / SRP-dependent cotranslational protein targeting to membrane, translocation / signal sequence binding / SRP-dependent cotranslational protein targeting to membrane / post-translational protein targeting to membrane, translocation / peptide transmembrane transporter activity / nuclear inner membrane / retrograde protein transport, ER to cytosol / protein transmembrane transporter activity / ERAD pathway / guanyl-nucleotide exchange factor activity / cell periphery / ribosome binding / endoplasmic reticulum membrane / structural molecule activity / endoplasmic reticulum / mitochondrion / membrane / cytosol Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.0 Å | |||||||||
![]() | Itskanov S / Park E | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Stepwise gating of the Sec61 protein-conducting channel by Sec63 and Sec62. Authors: Samuel Itskanov / Katie M Kuo / James C Gumbart / Eunyong Park / ![]() Abstract: Many proteins are transported into the endoplasmic reticulum by the universally conserved Sec61 channel. Post-translational transport requires two additional proteins, Sec62 and Sec63, but their ...Many proteins are transported into the endoplasmic reticulum by the universally conserved Sec61 channel. Post-translational transport requires two additional proteins, Sec62 and Sec63, but their functions are poorly defined. In the present study, we determined cryo-electron microscopy (cryo-EM) structures of several variants of Sec61-Sec62-Sec63 complexes from Saccharomyces cerevisiae and Thermomyces lanuginosus and show that Sec62 and Sec63 induce opening of the Sec61 channel. Without Sec62, the translocation pore of Sec61 remains closed by the plug domain, rendering the channel inactive. We further show that the lateral gate of Sec61 must first be partially opened by interactions between Sec61 and Sec63 in cytosolic and luminal domains, a simultaneous disruption of which completely closes the channel. The structures and molecular dynamics simulations suggest that Sec62 may also prevent lipids from invading the channel through the open lateral gate. Our study shows how Sec63 and Sec62 work together in a hierarchical manner to activate Sec61 for post-translational protein translocation. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 32.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 23.5 KB 23.5 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 9.2 KB | Display | ![]() |
Images | ![]() | 29.3 KB | ||
Masks | ![]() | 64 MB | ![]() | |
Others | ![]() ![]() ![]() | 59.7 MB 59.5 MB 59.5 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 482.3 KB | Display | ![]() |
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Full document | ![]() | 481.8 KB | Display | |
Data in XML | ![]() | 14.9 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7kahC ![]() 7kaiC ![]() 7kajC ![]() 7kakC ![]() 7kalC ![]() 7kamC ![]() 7kanC ![]() 7kaoC ![]() 7kapC ![]() 7kaqC ![]() 7karC ![]() 7kasC ![]() 7katC ![]() 7kauC ![]() 7kb5C C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Unsharpened, lowpass-filtered map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.19 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
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-Additional map: Sharpened map
File | emd_22785_additional_1.map | ||||||||||||
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Annotation | Sharpened map | ||||||||||||
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Density Histograms |
-Half map: half-volume 1
File | emd_22785_half_map_1.map | ||||||||||||
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Annotation | half-volume 1 | ||||||||||||
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-Half map: half-volume 2
File | emd_22785_half_map_2.map | ||||||||||||
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Annotation | half-volume 2 | ||||||||||||
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Density Histograms |
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Sample components
-Entire : Endoplasmic reticulum protein-transport machinery Sec complex fro...
Entire | Name: Endoplasmic reticulum protein-transport machinery Sec complex from yeast |
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Components |
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-Supramolecule #1: Endoplasmic reticulum protein-transport machinery Sec complex fro...
Supramolecule | Name: Endoplasmic reticulum protein-transport machinery Sec complex from yeast type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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-Macromolecule #1: Protein transport channel Sec61 complex, alpha subunit (Sec61)
Macromolecule | Name: Protein transport channel Sec61 complex, alpha subunit (Sec61) type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: MSSNRVLDLF KPFESFLPEV IAPERKVPYN QKLIWTGVSL LIFLILGQIP LYGIVSSETS DPLYWLRAML ASNRGTLLEL GVSPIITSS MIFQFLQGTQ LLQIRPESKQ DRELFQIAQK VCAIILILGQ ALVVVMTGNY GAPSDLGLPI CLLLIFQLMF A SLIVMLLD ...String: MSSNRVLDLF KPFESFLPEV IAPERKVPYN QKLIWTGVSL LIFLILGQIP LYGIVSSETS DPLYWLRAML ASNRGTLLEL GVSPIITSS MIFQFLQGTQ LLQIRPESKQ DRELFQIAQK VCAIILILGQ ALVVVMTGNY GAPSDLGLPI CLLLIFQLMF A SLIVMLLD ELLSKGYGLG SGISLFTATN IAEQIFWRAF APTTVNSGRG KEFEGAVIAF FHLLAVRKDK KRALVEAFYR TN LPNMFQV LMTVAIFLFV LYLQGFRYEL PIRSTKVRGQ IGIYPIKLFY TSNTPIMLQS ALTSNIFLIS QILFQKYPTN PLI RLIGVW GIRPGTQGPQ MALSGLAYYI QPLMSLSEAL LDPIKTIVYI TFVLGSCAVF SKTWIEISGT SPRDIAKQFK DQGM VINGK RETSIYRELK KIIPTAAAFG GATIGALSVG SDLLGTLGSG ASILMATTTI YGYYEAAAKE GGFTKNLVPG FSDLM |
-Macromolecule #2: Protein transport channel Sec61 complex, beta subunit (Sbh1)
Macromolecule | Name: Protein transport channel Sec61 complex, beta subunit (Sbh1) type: protein_or_peptide / ID: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: MSSPTPPGGQ RTLQKRKQGS SQKVAASAPK KNTNSNNSIL KIYSDEATGL RVDPLVVLFL AVGFIFSVVA LHVISKVAGK LF |
-Macromolecule #3: Protein transport channel Sec61 complex, gamma subunit (Sss1)
Macromolecule | Name: Protein transport channel Sec61 complex, gamma subunit (Sss1) type: protein_or_peptide / ID: 3 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: MARASEKGEE KKQSNNQVEK LVEAPVEFVR EGTQFLAKCK KPDLKEYTKI VKAVGIGFIA VGIIGYAIKL IHIPIRYVIV |
-Macromolecule #4: Protein transport protein Sec63
Macromolecule | Name: Protein transport protein Sec63 / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: GGSGGSGGSG GSGGSPTNYE YDEASETWPS FILTGLLMVV GPMTLLQIYQ IFFGANAEDG NSGKSKEFNE EVFKNLNEEY TSDEIKQFR RKFDKNSNKK SKIWSRRNII IIVGWILVAI LLQRINSNDA IKDAATKLFD PYEILGISTS ASDRDIKSAY R KLSVKFHP ...String: GGSGGSGGSG GSGGSPTNYE YDEASETWPS FILTGLLMVV GPMTLLQIYQ IFFGANAEDG NSGKSKEFNE EVFKNLNEEY TSDEIKQFR RKFDKNSNKK SKIWSRRNII IIVGWILVAI LLQRINSNDA IKDAATKLFD PYEILGISTS ASDRDIKSAY R KLSVKFHP DKLAKGLTPD EKSVMEETYV QITKAYESLT DELVRQNYLK YGHPDGPQST SHGIALPRFL VDGSASPLLV VC YVALLGL ILPYFVSRWW ARTQSYTKKG IHNVTASNFV SNLVNYKPSE IVTTDLILHW LSFAHEFKQF FPDLQPTDFE KLL QDHINR RDSGKLNNAK FRIVAKCHSL LHGLLDIACG FRNLDIALGA INTFKCIVQA VPLTPNCQIL QLPNVDKEHF ITKT GDIHT LGKLFTLEDA KIGEVLGIKD QAKLNETLRV ASHIPNLKII KADFLVPGEN QVTPSSTPYI SLKVLVRSAK QPLIP TSLI PEENLTEPQD FESQRDPFAM MSKQPLVPYS FAPFFPTKRR GSWCCLVSSQ KDGKILQTPI IIEKLSYKNL NDDKDF FDK RIKMDLTKHE KFDINDWEIG TIKIPLGQPA PETVGDFFFR VIVKSTDYFT TDLDITMNMK VRDSPAVEQV EVYSEED DE YSTDDDETES DDESDASDYT DIDTDTEAED DESPEAGGAT TASGTGENLY FQ |
-Macromolecule #5: Protein transport protein Sec66/Sec71
Macromolecule | Name: Protein transport protein Sec66/Sec71 / type: protein_or_peptide / ID: 5 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: MSEFNETKFS NNGTFFETEE PIVETKSISV YTPLIYVFIL VVSLVMFASS YRKKQAKKIS EQPSIFDEND AHDLYFQIKE MSENEKIHE KVLKAALLNR GAESVRRSLK LKELAPQINL LYKNGSIGED YWKRFETEVK LIELEFKDTL QEAERLQPGW V QLFVMVCK ...String: MSEFNETKFS NNGTFFETEE PIVETKSISV YTPLIYVFIL VVSLVMFASS YRKKQAKKIS EQPSIFDEND AHDLYFQIKE MSENEKIHE KVLKAALLNR GAESVRRSLK LKELAPQINL LYKNGSIGED YWKRFETEVK LIELEFKDTL QEAERLQPGW V QLFVMVCK EICFNQALSR RYQSILKRKE VCIKEWELKI NNDGRLVN |
-Macromolecule #6: Protein transport protein Sec72
Macromolecule | Name: Protein transport protein Sec72 / type: protein_or_peptide / ID: 6 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: MVTLEYNANS KLITASDAVV ALSTETNIDQ INVLTTSLIG ETNPNFTPQP NEALSKMIKG LFESGMKNLQ QKKLNEALKN VSLAIEMAQ RKRAPWEAFA IQLPELHFML RSKIDLCLIL GKHLEALQDL DFLLGTGLIQ PDVFVRKADC LLKLRQWEEA R ATCERGLA ...String: MVTLEYNANS KLITASDAVV ALSTETNIDQ INVLTTSLIG ETNPNFTPQP NEALSKMIKG LFESGMKNLQ QKKLNEALKN VSLAIEMAQ RKRAPWEAFA IQLPELHFML RSKIDLCLIL GKHLEALQDL DFLLGTGLIQ PDVFVRKADC LLKLRQWEEA R ATCERGLA LAPEDMKLRA LLIETARNLA EYNGE |
-Macromolecule #7: Protein transport protein Sec62
Macromolecule | Name: Protein transport protein Sec62 / type: protein_or_peptide / ID: 7 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: MSAVGPGSNA GASVNGGSAT AIATLLRNHK ELKQRQGLFQ AKQTDFFRYK RFVRALHSEE YANKSARQP EIYPTIPSNK IEDQLKSREI FIQLIKAQMV IPVKKLHSQE CKEHGLKPSK D FPHLIVSN KAQLEADEYF VWNYNPRTYM DYLIVIGVVS IILALVCYPL ...String: MSAVGPGSNA GASVNGGSAT AIATLLRNHK ELKQRQGLFQ AKQTDFFRYK RFVRALHSEE YANKSARQP EIYPTIPSNK IEDQLKSREI FIQLIKAQMV IPVKKLHSQE CKEHGLKPSK D FPHLIVSN KAQLEADEYF VWNYNPRTYM DYLIVIGVVS IILALVCYPL WPRSMRRGSY YV SLGAFGI LAGFFAVAIL RLILYVLSLI VYKDVGGFWI FPNLFEDCGV LESFKPLYGF GEK DTYSYK KKLKRMKKKQ AKRESNKKKA INEKAEQN |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 5 mg/mL |
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Buffer | pH: 7.5 |
Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.039 kPa |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Quantum LS / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 49.1 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 42017 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |