+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-0645 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Saccharomyces cerevisiae V-ATPase Stv1-VO | |||||||||
Map data | ||||||||||
Sample |
| |||||||||
Keywords | Proton pump / MEMBRANE PROTEIN | |||||||||
Function / homology | Function and homology information cell wall mannoprotein biosynthetic process / ATPase-coupled ion transmembrane transporter activity / Insulin receptor recycling / Transferrin endocytosis and recycling / ROS and RNS production in phagocytes / Amino acids regulate mTORC1 / Golgi lumen acidification / P-type proton-exporting transporter activity / vacuolar transport / vacuolar proton-transporting V-type ATPase, V0 domain ...cell wall mannoprotein biosynthetic process / ATPase-coupled ion transmembrane transporter activity / Insulin receptor recycling / Transferrin endocytosis and recycling / ROS and RNS production in phagocytes / Amino acids regulate mTORC1 / Golgi lumen acidification / P-type proton-exporting transporter activity / vacuolar transport / vacuolar proton-transporting V-type ATPase, V0 domain / endosomal lumen acidification / protein targeting to vacuole / vacuole organization / vacuolar proton-transporting V-type ATPase complex / proton-transporting V-type ATPase complex / fungal-type vacuole / vacuolar acidification / fungal-type vacuole membrane / phosphatidylinositol-4-phosphate binding / proton transmembrane transporter activity / intracellular copper ion homeostasis / Neutrophil degranulation / proton-transporting ATPase activity, rotational mechanism / RNA endonuclease activity / proton transmembrane transport / cell periphery / transmembrane transport / endocytosis / late endosome / ATPase binding / protein-containing complex assembly / intracellular iron ion homeostasis / endosome membrane / Golgi membrane / endoplasmic reticulum membrane / Golgi apparatus / membrane Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||
Authors | Vasanthakumar T / Bueler SA / Wu D / Beilsten-Edmands V / Robinson CV / Rubinstein JL | |||||||||
Funding support | Canada, 1 items
| |||||||||
Citation | Journal: Proc Natl Acad Sci U S A / Year: 2019 Title: Structural comparison of the vacuolar and Golgi V-ATPases from . Authors: Thamiya Vasanthakumar / Stephanie A Bueler / Di Wu / Victoria Beilsten-Edmands / Carol V Robinson / John L Rubinstein / Abstract: Proton-translocating vacuolar-type ATPases (V-ATPases) are necessary for numerous processes in eukaryotic cells, including receptor-mediated endocytosis, protein maturation, and lysosomal ...Proton-translocating vacuolar-type ATPases (V-ATPases) are necessary for numerous processes in eukaryotic cells, including receptor-mediated endocytosis, protein maturation, and lysosomal acidification. In mammals, V-ATPase subunit isoforms are differentially targeted to various intracellular compartments or tissues, but how these subunit isoforms influence enzyme activity is not clear. In the yeast , isoform diversity is limited to two different versions of the proton-translocating subunit a: Vph1p, which is targeted to the vacuole, and Stv1p, which is targeted to the Golgi apparatus and endosomes. We show that purified V-ATPase complexes containing Vph1p have higher ATPase activity than complexes containing Stv1p and that the relative difference in activity depends on the presence of lipids. We also show that V complexes containing Stv1p could be readily purified without attached V regions. We used this effect to determine structures of the membrane-embedded V region with Stv1p at 3.1-Å resolution, which we compare with a structure of the V region with Vph1p that we determine to 3.2-Å resolution. These maps reveal differences in the surface charge near the cytoplasmic proton half-channel. Both maps also show the presence of bound lipids, as well as regularly spaced densities that may correspond to ergosterol or bound detergent, around the c-ring. | |||||||||
History |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_0645.map.gz | 59.3 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-0645-v30.xml emd-0645.xml | 18.6 KB 18.6 KB | Display Display | EMDB header |
Images | emd_0645.png | 39.5 KB | ||
Filedesc metadata | emd-0645.cif.gz | 6.8 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-0645 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-0645 | HTTPS FTP |
-Validation report
Summary document | emd_0645_validation.pdf.gz | 547.6 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_0645_full_validation.pdf.gz | 547.2 KB | Display | |
Data in XML | emd_0645_validation.xml.gz | 6.5 KB | Display | |
Data in CIF | emd_0645_validation.cif.gz | 7.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-0645 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-0645 | HTTPS FTP |
-Related structure data
Related structure data | 6o7uMC 0644C 0646C 0647C 0648C 6o7tC 6o7vC 6o7wC 6o7xC C: citing same article (ref.) M: atomic model generated by this map |
---|---|
Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_0645.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.06 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Sample components
-Entire : Saccharomyces cerevisiae V-ATPase Stv1-VO
Entire | Name: Saccharomyces cerevisiae V-ATPase Stv1-VO |
---|---|
Components |
|
-Supramolecule #1: Saccharomyces cerevisiae V-ATPase Stv1-VO
Supramolecule | Name: Saccharomyces cerevisiae V-ATPase Stv1-VO / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
-Macromolecule #1: V-type proton ATPase subunit a, Golgi isoform
Macromolecule | Name: V-type proton ATPase subunit a, Golgi isoform / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Molecular weight | Theoretical: 104.481141 KDa |
Sequence | String: MNQEEAIFRS ADMTYVQLYI PLEVIREVTF LLGKMSVFMV MDLNKDLTAF QRGYVNQLRR FDEVERMVGF LNEVVEKHAA ETWKYILHI DDEGNDIAQP DMADLINTME PLSLENVNDM VKEITDCESR ARQLDESLDS LRSKLNDLLE QRQVIFECSK F IEVNPGIA ...String: MNQEEAIFRS ADMTYVQLYI PLEVIREVTF LLGKMSVFMV MDLNKDLTAF QRGYVNQLRR FDEVERMVGF LNEVVEKHAA ETWKYILHI DDEGNDIAQP DMADLINTME PLSLENVNDM VKEITDCESR ARQLDESLDS LRSKLNDLLE QRQVIFECSK F IEVNPGIA GRATNPEIEQ EERDVDEFRM TPDDISETLS DAFSFDDETP QDRGALGNDL TRNQSVEDLS FLEQGYQHRY MI TGSIRRT KVDILNRILW RLLRGNLIFQ NFPIEEPLLE GKEKVEKDCF IIFTHGETLL KKVKRVIDSL NGKIVSLNTR SSE LVDTLN RQIDDLQRIL DTTEQTLHTE LLVIHDQLPV WSAMTKREKY VYTTLNKFQQ ESQGLIAEGW VPSTELIHLQ DSLK DYIET LGSEYSTVFN VILTNKLPPT YHRTNKFTQA FQSIVDAYGI ATYKEINAGL ATVVTFPFMF AIMFGDMGHG FILFL MALF LVLNERKFGA MHRDEIFDMA FTGRYVLLLM GAFSVYTGLL YNDIFSKSMT IFKSGWQWPS TFRKGESIEA KKTGVY PFG LDFAWHGTDN GLLFSNSYKM KLSILMGYAH MTYSFMFSYI NYRAKNSKVD IIGNFIPGLV FMQSIFGYLS WAIVYKW SK DWIKDDKPAP GLLNMLINMF LAPGTIDDQL YSGQAKLQVV LLLAALVCVP WLLLYKPLTL RRLNKNGGGG RPHGYQSV G NIEHEEQIAQ QRHSAEGFQG MIISDVASVA DSINESVGGG EQGPFNFGDV MIHQVIHTIE FCLNCISHTA SYLRLWALS LAHAQLSSVL WDMTISNAFS SKNSGSPLAV MKVVFLFAMW FVLTVCILVF MEGTSAMLHA LRLHWVEAMS KFFEGEGYAY EPFSFRAII EDYKDHDGDY KDHDIDYKDD DDK UniProtKB: V-type proton ATPase subunit a, Golgi isoform |
-Macromolecule #2: V0 assembly protein 1
Macromolecule | Name: V0 assembly protein 1 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Molecular weight | Theoretical: 29.694885 KDa |
Sequence | String: MVFGQLYALF IFTLSCCISK TVQADSSKES SSFISFDKES NWDTISTISS TADVISSVDS AIAVFEFDNF SLLDNLMIDE EYPFFNRFF ANDVSLTVHD DSPLNISQSL SPIMEQFTVD ELPESASDLL YEYSLDDKSI VLFKFTSDAY DLKKLDEFID S CLSFLEDK ...String: MVFGQLYALF IFTLSCCISK TVQADSSKES SSFISFDKES NWDTISTISS TADVISSVDS AIAVFEFDNF SLLDNLMIDE EYPFFNRFF ANDVSLTVHD DSPLNISQSL SPIMEQFTVD ELPESASDLL YEYSLDDKSI VLFKFTSDAY DLKKLDEFID S CLSFLEDK SGDNLTVVIN SLGWAFEDED GDDEYATEET LSHHDNNKGK EGDDDILSSI WTEGLLMCLI VSALLLFILI VA LSWISNL DITYGALEKS TNPIKKNN UniProtKB: V0 assembly protein 1 |
-Macromolecule #3: V-type proton ATPase subunit c''
Macromolecule | Name: V-type proton ATPase subunit c'' / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Molecular weight | Theoretical: 22.610641 KDa |
Sequence | String: MNKESKDDDM SLGKFSFSHF LYYLVLIVVI VYGLYKLFTG HGSDINFGKF LLRTSPYMWA NLGIALCVGL SVVGAAWGIF ITGSSMIGA GVRAPRITTK NLISIIFCEV VAIYGLIIAI VFSSKLTVAT AENMYSKSNL YTGYSLFWAG ITVGASNLIC G IAVGITGA ...String: MNKESKDDDM SLGKFSFSHF LYYLVLIVVI VYGLYKLFTG HGSDINFGKF LLRTSPYMWA NLGIALCVGL SVVGAAWGIF ITGSSMIGA GVRAPRITTK NLISIIFCEV VAIYGLIIAI VFSSKLTVAT AENMYSKSNL YTGYSLFWAG ITVGASNLIC G IAVGITGA TAAISDAADS ALFVKILVIE IFGSILGLLG LIVGLLMAGK ASEFQ UniProtKB: V-type proton ATPase subunit c'' |
-Macromolecule #4: V-type proton ATPase subunit d
Macromolecule | Name: V-type proton ATPase subunit d / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Molecular weight | Theoretical: 39.822484 KDa |
Sequence | String: MEGVYFNIDN GFIEGVVRGY RNGLLSNNQY INLTQCDTLE DLKLQLSSTD YGNFLSSVSS ESLTTSLIQE YASSKLYHEF NYIRDQSSG STRKFMDYIT YGYMIDNVAL MITGTIHDRD KGEILQRCHP LGWFDTLPTL SVATDLESLY ETVLVDTPLA P YFKNCFDT ...String: MEGVYFNIDN GFIEGVVRGY RNGLLSNNQY INLTQCDTLE DLKLQLSSTD YGNFLSSVSS ESLTTSLIQE YASSKLYHEF NYIRDQSSG STRKFMDYIT YGYMIDNVAL MITGTIHDRD KGEILQRCHP LGWFDTLPTL SVATDLESLY ETVLVDTPLA P YFKNCFDT AEELDDMNIE IIRNKLYKAY LEDFYNFVTE EIPEPAKECM QTLLGFEADR RSINIALNSL QSSDIDPDLK SD LLPNIGK LYPLATFHLA QAQDFEGVRA ALANVYEYRG FLETGNLEDH FYQLEMELCR DAFTQQFAIS TVWAWMKSKE QEV RNITWI AECIAQNQRE RINNYISVY UniProtKB: V-type proton ATPase subunit d |
-Macromolecule #5: V-type proton ATPase subunit e
Macromolecule | Name: V-type proton ATPase subunit e / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Molecular weight | Theoretical: 8.387065 KDa |
Sequence | String: MSSFYTVVGV FIVVSAMSVL FWIMAPKNNQ AVWRSTVILT LAMMFLMWAI TFLCQLHPLV APRRSDLRPE FAE UniProtKB: V-type proton ATPase subunit e |
-Macromolecule #6: Putative protein YPR170W-B
Macromolecule | Name: Putative protein YPR170W-B / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Molecular weight | Theoretical: 9.369934 KDa |
Sequence | String: MRPVVSTGKA WCCTVLSAFG VVILSVIAHL FNTNHESFVG SINDPEDGPA VAHTVYLAAL VYLVFFVFCG FQVYLARRKP SIELR UniProtKB: V-type proton ATPase subunit f |
-Macromolecule #7: V-type proton ATPase subunit c
Macromolecule | Name: V-type proton ATPase subunit c / type: protein_or_peptide / ID: 7 / Number of copies: 8 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Molecular weight | Theoretical: 16.357501 KDa |
Sequence | String: MTELCPVYAP FFGAIGCASA IIFTSLGAAY GTAKSGVGIC ATCVLRPDLL FKNIVPVIMA GIIAIYGLVV SVLVCYSLGQ KQALYTGFI QLGAGLSVGL SGLAAGFAIG IVGDAGVRGS SQQPRLFVGM ILILIFAEVL GLYGLIVALL LNSRATQDVV C UniProtKB: V-type proton ATPase subunit c |
-Macromolecule #8: V-type proton ATPase subunit c'
Macromolecule | Name: V-type proton ATPase subunit c' / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Molecular weight | Theoretical: 17.046361 KDa |
Sequence | String: MSTQLASNIY APLYAPFFGF AGCAAAMVLS CLGAAIGTAK SGIGIAGIGT FKPELIMKSL IPVVMSGILA IYGLVVAVLI AGNLSPTED YTLFNGFMHL SCGLCVGFAC LSSGYAIGMV GDVGVRKYMH QPRLFVGIVL ILIFSEVLGL YGMIVALILN T RGSE UniProtKB: V-type proton ATPase subunit c' |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
---|---|
Vitrification | Cryogen name: ETHANE-PROPANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Average electron dose: 42.7 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: OTHER / Details: Ab initio |
---|---|
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 163024 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |