[English] 日本語
Yorodumi- EMDB-20875: Upper fiber-focused reconstruction of the Staphylococcus aureus p... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-20875 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Upper fiber-focused reconstruction of the Staphylococcus aureus phage 80alpha baseplate | |||||||||
Map data | C6, full particle reconstruction following C6, FibU-focused 3D classification of non-FibU signal-subtracted particles. Locally autosharpened in Phenix and used for final FibU model refinement. | |||||||||
Sample |
| |||||||||
Function / homology | Function and homology information | |||||||||
Biological species | Staphylococcus virus 80alpha | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.14 Å | |||||||||
Authors | Kizziah JL / Dokland T | |||||||||
Funding support | United States, 1 items
| |||||||||
Citation | Journal: PLoS Pathog / Year: 2020 Title: Structure of the host cell recognition and penetration machinery of a Staphylococcus aureus bacteriophage. Authors: James L Kizziah / Keith A Manning / Altaira D Dearborn / Terje Dokland / Abstract: Staphylococcus aureus is a common cause of infections in humans. The emergence of virulent, antibiotic-resistant strains of S. aureus is a significant public health concern. Most virulence and ...Staphylococcus aureus is a common cause of infections in humans. The emergence of virulent, antibiotic-resistant strains of S. aureus is a significant public health concern. Most virulence and resistance factors in S. aureus are encoded by mobile genetic elements, and transduction by bacteriophages represents the main mechanism for horizontal gene transfer. The baseplate is a specialized structure at the tip of bacteriophage tails that plays key roles in host recognition, cell wall penetration, and DNA ejection. We have used high-resolution cryo-electron microscopy to determine the structure of the S. aureus bacteriophage 80α baseplate at 3.75 Å resolution, allowing atomic models to be built for most of the major tail and baseplate proteins, including two tail fibers, the receptor binding protein, and part of the tape measure protein. Our structure provides a structural basis for understanding host recognition, cell wall penetration and DNA ejection in viruses infecting Gram-positive bacteria. Comparison to other phages demonstrates the modular design of baseplate proteins, and the adaptations to the host that take place during the evolution of staphylococci and other pathogens. | |||||||||
History |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_20875.map.gz | 469.4 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-20875-v30.xml emd-20875.xml | 22.6 KB 22.6 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_20875_fsc.xml | 18.1 KB | Display | FSC data file |
Images | emd_20875.png | 52.7 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-20875 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-20875 | HTTPS FTP |
-Validation report
Summary document | emd_20875_validation.pdf.gz | 379 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_20875_full_validation.pdf.gz | 378.5 KB | Display | |
Data in XML | emd_20875_validation.xml.gz | 16.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20875 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20875 | HTTPS FTP |
-Related structure data
Related structure data | 6v8iMC C: citing same article (ref.) M: atomic model generated by this map |
---|---|
Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|
-Map
File | Download / File: emd_20875.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | C6, full particle reconstruction following C6, FibU-focused 3D classification of non-FibU signal-subtracted particles. Locally autosharpened in Phenix and used for final FibU model refinement. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.21 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Sample components
-Entire : Staphylococcus phage 80alpha baseplate
Entire | Name: Staphylococcus phage 80alpha baseplate |
---|---|
Components |
|
-Supramolecule #1: Staphylococcus phage 80alpha baseplate
Supramolecule | Name: Staphylococcus phage 80alpha baseplate / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all Details: 80alpha baseplate attached to fully formed tails with no capsids |
---|---|
Source (natural) | Organism: Staphylococcus virus 80alpha / Strain: ST247 |
Recombinant expression | Organism: Staphylococcus aureus (bacteria) |
Molecular weight | Theoretical: 3.7 MDa |
-Macromolecule #1: Major Tail Protein
Macromolecule | Name: Major Tail Protein / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Staphylococcus virus 80alpha |
Recombinant expression | Organism: Staphylococcus aureus (bacteria) |
Sequence | String: MANMKNSNDR IILFRKAGEK VDATKMLFLT EYGLSHEADT DTEDTMDGSY NTGGSVESTM SGTAKMFYGD DFADEIEDAV VDRVLYEAWE VESRIPGKNG DSAKFKAKYF QGFHNKFELK AEANGIDEYE YEYGVNGRFQ RGFATLPEAV TKKLKATGYR FHDTTKEDAL TSEDLTAIPQ PKVDSSTVTP GEV |
-Macromolecule #2: Distal Tail Protein
Macromolecule | Name: Distal Tail Protein / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Staphylococcus virus 80alpha |
Recombinant expression | Organism: Staphylococcus aureus (bacteria) |
Sequence | String: MDIELTKKDG TVIKLSEYGF IVNDIVIDSM QINTKYQDKE NMNGRILMGS NYISRDIVVP CFCKVKNRSD IAYMRDMLYS LTTDIEPMYL REIRRKEELN YRFTQPTSDD YVKLDKNNFP DYEYSRHDQQ IYVNGKQYKV IFNGVINPKQ KGNKVSFELK FETTELPYGE ...String: MDIELTKKDG TVIKLSEYGF IVNDIVIDSM QINTKYQDKE NMNGRILMGS NYISRDIVVP CFCKVKNRSD IAYMRDMLYS LTTDIEPMYL REIRRKEELN YRFTQPTSDD YVKLDKNNFP DYEYSRHDQQ IYVNGKQYKV IFNGVINPKQ KGNKVSFELK FETTELPYGE SIGTSLELEE NKKVGLWSFD FNIDWHAGGD KRKYTFENLS KGTVYYHGSA PNDQFNMYKK ITIILGEDTE SFVWNLTHAE IMKIEGIKLK AGDRIVYDSF RVYKNGVEIS TETNISQPKF KYGANKFEFN QTVQKVQFDL KFYYK |
-Macromolecule #3: Tail-Associated Lysin
Macromolecule | Name: Tail-Associated Lysin / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Staphylococcus virus 80alpha |
Recombinant expression | Organism: Staphylococcus aureus (bacteria) |
Sequence | String: MTITIKPPKG NGAPVPVETT LVKKVNADGV LTFDILENKY TYEVINAIGK RWIVSHVEGE NDKKEYVITV IDRKSEGDRQ LVECTAREIP IDKLMIDRIY VNVTGSFTVE RYFNIVFQGT GMLFEVEGKV KSSKFENGGE GDTRLEMFKK GLEHFGLEYK ITYDKKKDRY ...String: MTITIKPPKG NGAPVPVETT LVKKVNADGV LTFDILENKY TYEVINAIGK RWIVSHVEGE NDKKEYVITV IDRKSEGDRQ LVECTAREIP IDKLMIDRIY VNVTGSFTVE RYFNIVFQGT GMLFEVEGKV KSSKFENGGE GDTRLEMFKK GLEHFGLEYK ITYDKKKDRY KFVLTPFANQ KASYFISDEV NANAIKLEED ASDFATFIRG YGNYSGEETF EHAGLVMEAR SALAEIYGDI HAEPFKDGKV TDQETMDKEL QSRLKKSLKQ SLSLDFLVLR ESYPEADPQP GDIVQIKSTK LGLNDLVRIV QVKTIRGINN VIVKQDVTLG EFNREQRYMK KVNTAANYVS GLNDVNLSNP SKAAENLKSK VASIAKSTLD LMSRTDLIED KQQKVSSKTV TTSDGTIVHD FIDKSNIKDV KTIGTIGDSV ARGSHAKTNF TEMLGKKLKA KTTNLARGGA TMATVPIGKE AVENSIYRQA EQIRGDLIIL QGTDDDWLHG YWAGVPIGTD KTDTKTFYGA FCSAIEVIRK NNPASKILVM TATRQCPMSG TMIRRKDTDK NKLGLTLEDY VNAQILACSE LDVPVYDAYH TDYFKPYNPA FRKSSMPDGL HPNERGHEVI MYELIKNYYQ FYG |
-Macromolecule #4: Receptor Binding Protein
Macromolecule | Name: Receptor Binding Protein / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Staphylococcus virus 80alpha |
Recombinant expression | Organism: Staphylococcus aureus (bacteria) |
Sequence | String: MDNKLITDLS RVFDYRYVDE NEYNFKLISD MLTDFNFSLE YHRNKEVFAH NGEQIKYEHL NVTSSVSDFL TYLNGRFSNM VLGHNGDGIN EVKDARVDNT GYDHKTLQDR LYHDYSTLDA FTKKVEKAVD ENYKEYRATE YRFEPKEQEP EFITDLSPYT NAVMQSFWVD ...String: MDNKLITDLS RVFDYRYVDE NEYNFKLISD MLTDFNFSLE YHRNKEVFAH NGEQIKYEHL NVTSSVSDFL TYLNGRFSNM VLGHNGDGIN EVKDARVDNT GYDHKTLQDR LYHDYSTLDA FTKKVEKAVD ENYKEYRATE YRFEPKEQEP EFITDLSPYT NAVMQSFWVD PRTKIIYMTQ ARPGNHYMLS RLKPNGQFID RLLVKNGGHG THNAYRYIGN ELWIYSAVLD ANENNKFVRF QYRTGEITYG NEMQDVMPNI FNDRYTSAIY NPIENLMIFR REYKASERQL KNSLNFVEVR SADDIDKGID KVLYQMDIPM EYTSDTQPMQ GITYDAGILY WYTGDSKPAN PNYLQGFDIK TKELLFKRRI DIGGVNNNFK GDFQEAEGLD MYYDLETGRK ALLIGVTIGP GNNRHHSIYS IGQRGVNQFL KNIAPQVSMT DSGGRVKPLP IQNPAYLSDI TEVGHYYIYT QDTQNALDFP LPKAFRDAGW FFDVLPGHYN GALRQVLTRN STGRNMLKFE RVIDIFNKKN NGAWNFCPQN AGYWEHIPKS ITKLSDLKIV GLDFYITTEE SNRFTDFPKD FKGIAGWILE VKSNTPGNTT QVLRRNNFPS AHQFLVRNFG TGGVGKWSLF EGKVVE |
-Macromolecule #5: Fiber Lower
Macromolecule | Name: Fiber Lower / type: protein_or_peptide / ID: 5 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Staphylococcus virus 80alpha |
Recombinant expression | Organism: Staphylococcus aureus (bacteria) |
Sequence | String: MVVDNFSKDD NLIELQTTSQ YNPVIDTNIS FYESDRGTGV LNFAVTKNNR PLSISSEHVK TFIVLKTDDY NVDRGAYISD ELTIVDAING RLQYVIPNEF LKHSGKVHAQ AFFTQNGSDN VVVERQFSFN IENDLVSGFD GITKLVYIKS IQDTIEAVGK DFNQLKQNMA ...String: MVVDNFSKDD NLIELQTTSQ YNPVIDTNIS FYESDRGTGV LNFAVTKNNR PLSISSEHVK TFIVLKTDDY NVDRGAYISD ELTIVDAING RLQYVIPNEF LKHSGKVHAQ AFFTQNGSDN VVVERQFSFN IENDLVSGFD GITKLVYIKS IQDTIEAVGK DFNQLKQNMA DTQTLIAKVN DSATKGIQQI EIKQNEAIQA ITATQTSATQ AVTAEFDKIV EKEQAIFERV NEVEQQINGA DLVKGNSTTN WQKSKLTDDY GKAIESYEQS IDSVLSAVNT SRIIHITSAT DAPSFKDIGT VDTPKEDGVD DGSDIPVAPN TLGKSGVLVV YVVDDSTARA TWYPDDSNDE YTKYKISGTW YPFYKKNDGD LTKQFVEETS NNALNQAKQY VDDKFGTTSW QQHKMTEANG QSIQVNLNNA QGDLGYLTAG NYYATRVPDL PGSVESYEGY LSVFVKDDTN KLFNFTPYNS KKIYTRSITN GRLEQQWTVP NEHKSTVLFD GGANGVGTTI NLTEPYTNYS ILLVSGTYPG GVIEGFGLTA LPNAIQLSKA NVVDSDGNGG GIYECLLSKT SSTTLRIDND VYFDLGKTSG SGANANKVTI TKIMGWK |
-Macromolecule #6: Fiber Upper
Macromolecule | Name: Fiber Upper / type: protein_or_peptide / ID: 6 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Staphylococcus virus 80alpha |
Recombinant expression | Organism: Staphylococcus aureus (bacteria) |
Sequence | String: MYKIKDVETR IKNDGVDLGD IGCRFYTEDE NTASIRIGIN DKQGRIDLKA HGLTPRLHLF MEDGSIFKNE PLIIDDVVKG FITYKIPKKV IKHAGYVRCK LFLEKEEEKI HVANFSFNIV DSGIESAVAK EIDVKLVDDA ITRILKDNAT DLLSKDFKEK IDKDVISYIE ...String: MYKIKDVETR IKNDGVDLGD IGCRFYTEDE NTASIRIGIN DKQGRIDLKA HGLTPRLHLF MEDGSIFKNE PLIIDDVVKG FITYKIPKKV IKHAGYVRCK LFLEKEEEKI HVANFSFNIV DSGIESAVAK EIDVKLVDDA ITRILKDNAT DLLSKDFKEK IDKDVISYIE KNESRFKGAK GDKGEPGQPG AKGEAGKKGE QGAPGKNGTV VSINPDTKMW QIDGKDTDIK AEPELLDKIN IANVEGLEDK LQEVEKIKDT TLNDSKTYTD TKIAELVDSA PESMNTLREL AEAIQNNSIS ESVLQQIGSK VSTEDFEEFK QTLNDLYAPK NHNHDERYVL SSQAFTKQQA DSLYQLKSAS QPTVKIWTGT ENEYNYIYQK DPNTLYLIKG |
-Macromolecule #7: Tape Measure Protein
Macromolecule | Name: Tape Measure Protein / type: protein_or_peptide / ID: 7 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Staphylococcus virus 80alpha |
Recombinant expression | Organism: Staphylococcus aureus (bacteria) |
Sequence | String: MTEYKIKATI EASVAKFKRQ IDSAVKSVQR FKRVADQTKD VELNANDKKL QKTIKVAKKS LDAFSNKNVK AKLDASIQDL QQKILESNFE LDKLNSKEAS PEVKLQKQKL TKDIAEAENK LSELEKKRVN IDVNADNSKF NRVLKVSKAS LEALNRSKAK AILDVDNSVA ...String: MTEYKIKATI EASVAKFKRQ IDSAVKSVQR FKRVADQTKD VELNANDKKL QKTIKVAKKS LDAFSNKNVK AKLDASIQDL QQKILESNFE LDKLNSKEAS PEVKLQKQKL TKDIAEAENK LSELEKKRVN IDVNADNSKF NRVLKVSKAS LEALNRSKAK AILDVDNSVA NSKIKRTKEE LKSIPNKTRS RLDVDTRLSI PTIYAFKKSL DALPNKKTTK VDVDTNGLKK VYAYIIKAND NFQRQMGNLA NMFRVFGTVG SNMVGGLLTS SFSILIPVIA SVVPVVFALL NAIKVLTGGV LALGGAVAIA GAGFVAFGAM AISAIKMLND GTLQASSATN EYKKALDGVK SAWTDIIKQN QSAIFTTLAN GLNTVKTAMQ SLQPFFSGIS RGMEEASQSV LKWAENSSVA SRFFNMMNTT GVSVFNKLLS AAGGFGDGLV NVFTQLAPLF QWSADWLDRL GQSFSNWANS AAGENSITRF IEYTKTNLPI IGNIFKNVFA GINNLMNAFS GSSTGIFQSL EQMTAKFREW SEQVGQSQGF KDFVSYIQTN GPLIMQLIGN IARGLVAFAT AMAPIASAVL RVAVAITGWI ANLFEAHPAT AQLVGVIITL VGAFRFLIAP ILAVMDFLGP LAARLVALVT KFGWAKTGTL VLSKAMTSLK GPIKLVTAIF QLLFGKIGLI RNAITGLVTV FGILGGPITI VIGVIAALIA IFVLLWNKNE GFRNFIINAW NAIKTFMVNV WNVLKAVASV VWNAILTAIT TAVSNVYNFI MIVWNQIVAY LQGLWNGIIA IATTVWNLLV TIITTVFTTI MTIVMTIWTA IWTFLSTIWN TIITIATTIW NLLVTVITTV FTTIMTIAMT IWNAIWTFLQ TLWNTIVTVA TKVWNAITTA ISTALQAAWS FISNIWNTIW SFLSGILTTI WNKVVSIFTQ VVSTISDKMS QAWNFIVTKG MQWVSTITST LINFVNRVIQ GFVNVVNKVS QGMTNAVNKI KSFIGDFVSA GADMIRGLIR GIGQMAGQLV DAAKNVAKKA LDAAKSALGI HSPSREFMDV GMYSMLGFVK GIDNHSSKVI RNVSNVADKV VDAFQPTLNA PDISSITGNL SNLGGNINAQ VQHTHSIETS PNMKTVKVEF DVNNDALTSI VNGRNAKRNS EYYL |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 Component:
| |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Grid | Model: Quantifoil R2/1 / Material: NICKEL / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV | |||||||||||||||
Details | 80alpha tails (with baseplates) purified by centrifugation, polyethylene glycol precipitation, and CsCl density gradation |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: DIRECT ELECTRON DE-20 (5k x 3k) / Digitization - Frames/image: 1-37 / Number grids imaged: 10 / Number real images: 6483 / Average exposure time: 1.156 sec. / Average electron dose: 104.33 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.7 µm / Nominal defocus min: 1.7 µm / Nominal magnification: 29000 |
Sample stage | Specimen holder model: GATAN 626 SINGLE TILT LIQUID NITROGEN CRYO TRANSFER HOLDER Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |