+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-11277 | ||||||||||||||||||
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Title | Open-closed state of the Bt1762-Bt1763 levan transport system | ||||||||||||||||||
Map data | Map filtered by local resolution using RELION's own implementation | ||||||||||||||||||
Sample |
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Function / homology | Function and homology information | ||||||||||||||||||
Biological species | Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482) (bacteria) | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.7 Å | ||||||||||||||||||
Authors | White JBR / van den Berg B / Ranson NA | ||||||||||||||||||
Funding support | United Kingdom, Switzerland, 5 items
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Citation | Journal: Nat Commun / Year: 2021 Title: Insights into SusCD-mediated glycan import by a prominent gut symbiont. Authors: Declan A Gray / Joshua B R White / Abraham O Oluwole / Parthasarathi Rath / Amy J Glenwright / Adam Mazur / Michael Zahn / Arnaud Baslé / Carl Morland / Sasha L Evans / Alan Cartmell / ...Authors: Declan A Gray / Joshua B R White / Abraham O Oluwole / Parthasarathi Rath / Amy J Glenwright / Adam Mazur / Michael Zahn / Arnaud Baslé / Carl Morland / Sasha L Evans / Alan Cartmell / Carol V Robinson / Sebastian Hiller / Neil A Ranson / David N Bolam / Bert van den Berg / Abstract: In Bacteroidetes, one of the dominant phyla of the mammalian gut, active uptake of large nutrients across the outer membrane is mediated by SusCD protein complexes via a "pedal bin" transport ...In Bacteroidetes, one of the dominant phyla of the mammalian gut, active uptake of large nutrients across the outer membrane is mediated by SusCD protein complexes via a "pedal bin" transport mechanism. However, many features of SusCD function in glycan uptake remain unclear, including ligand binding, the role of the SusD lid and the size limit for substrate transport. Here we characterise the β2,6 fructo-oligosaccharide (FOS) importing SusCD from Bacteroides thetaiotaomicron (Bt1762-Bt1763) to shed light on SusCD function. Co-crystal structures reveal residues involved in glycan recognition and suggest that the large binding cavity can accommodate several substrate molecules, each up to ~2.5 kDa in size, a finding supported by native mass spectrometry and isothermal titration calorimetry. Mutational studies in vivo provide functional insights into the key structural features of the SusCD apparatus and cryo-EM of the intact dimeric SusCD complex reveals several distinct states of the transporter, directly visualising the dynamics of the pedal bin transport mechanism. | ||||||||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_11277.map.gz | 42.8 MB | EMDB map data format | |
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Header (meta data) | emd-11277-v30.xml emd-11277.xml | 20.8 KB 20.8 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_11277_fsc.xml | 9.5 KB | Display | FSC data file |
Images | emd_11277.png | 18.5 KB | ||
Masks | emd_11277_msk_1.map | 70.2 MB | Mask map | |
Others | emd_11277_additional_1.map.gz emd_11277_additional_2.map.gz | 7.1 MB 54.3 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-11277 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-11277 | HTTPS FTP |
-Related structure data
Related structure data | 6zm1MC 6ytcC 6z8iC 6z9aC 6zazC 6zltC 6zluC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_11277.map.gz / Format: CCP4 / Size: 70.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Map filtered by local resolution using RELION's own implementation | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.07 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_11277_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: Sharpened post-processed map from RELION
File | emd_11277_additional_1.map | ||||||||||||
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Annotation | Sharpened post-processed map from RELION | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Unsharpened map from 3D auto-refine in RELION
File | emd_11277_additional_2.map | ||||||||||||
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Annotation | Unsharpened map from 3D auto-refine in RELION | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Dimeric Bt1762-Bt1763 levan transporter complex
Entire | Name: Dimeric Bt1762-Bt1763 levan transporter complex |
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Components |
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-Supramolecule #1: Dimeric Bt1762-Bt1763 levan transporter complex
Supramolecule | Name: Dimeric Bt1762-Bt1763 levan transporter complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Molecular weight | Theoretical: 360 KDa |
-Supramolecule #2: SusD homolog
Supramolecule | Name: SusD homolog / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482) (bacteria) |
Recombinant expression | Organism: Bacteroides thetaiotaomicron (bacteria) |
-Supramolecule #3: SusC homolog
Supramolecule | Name: SusC homolog / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2 |
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Source (natural) | Organism: Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482) (bacteria) |
-Macromolecule #1: SusD homolog
Macromolecule | Name: SusD homolog / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482) (bacteria) |
Molecular weight | Theoretical: 66.142625 KDa |
Recombinant expression | Organism: Bacteroides thetaiotaomicron (bacteria) |
Sequence | String: MKKIIYIATI GITLLTTSCD DFLDRQVPQG IVTGDQIASP EYVDNLVISA YAIWATGDDI NSSFSLWNYD VRSDDCYKGG SGTEDGGVF NALEISKGIN TTDWNINDIW KRLYQCITRA NTALQSLDQM DEKTYPLKNQ RIAEMRFLRG HAHFMLKQLF K KIVIVNDE ...String: MKKIIYIATI GITLLTTSCD DFLDRQVPQG IVTGDQIASP EYVDNLVISA YAIWATGDDI NSSFSLWNYD VRSDDCYKGG SGTEDGGVF NALEISKGIN TTDWNINDIW KRLYQCITRA NTALQSLDQM DEKTYPLKNQ RIAEMRFLRG HAHFMLKQLF K KIVIVNDE NMEPDAYNEL SNTTYTNDEQ WQKIADDFQF AYDNLPEVQI EKGRPAQAAA AAYLAKTYLY KAYRQDGADN AL TGINEED LKQVVKYTDP LIMAKGGYGL ETDYSMNFLP QYENGAESVW AIQYSINDGT YNGNLNWGMG LTTPQILGCC DFH KPSQNL VNAFKTDSQG KPLFSTYDNE NYEVATDNVD PRLFHTVGMP GFPYKYNEGY IIQKNDDWSR SKGLYGYYVS LKEN VDPDC DCLKKGSYWA SSLNHIVIRY ADVLLMRAEA LIQLNDGRIT DAISLINEVR SRAAGSTMLI FNYKEDYGVN FKVTP YDLK AYAQDEAMKM LKWERRVEFG MESSRFFDLV RWGEAKDVIN AYYVTEASRC SIYKNAGFTE NKNEYLPVPF EQISAS NGN YTQNFGWAAA AHHHHHH |
-Macromolecule #2: SusC homolog
Macromolecule | Name: SusC homolog / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482) (bacteria) |
Molecular weight | Theoretical: 115.483602 KDa |
Sequence | String: MPGIMKNKKL LCSVCFLFAF MSALWGQNIT VKGNVTSKTD GQPIIGASVV ETTATTNGTI TDFDGNFTLS VPVNSTLKIT YIGYKPVTV KAAAIVNVLL EEDTQMVDEV VVTGYTTQRK ADLTGAVSVV KVDEIQKQGE NNPVKALQGR VPGMNITADG N PSGSATVR ...String: MPGIMKNKKL LCSVCFLFAF MSALWGQNIT VKGNVTSKTD GQPIIGASVV ETTATTNGTI TDFDGNFTLS VPVNSTLKIT YIGYKPVTV KAAAIVNVLL EEDTQMVDEV VVTGYTTQRK ADLTGAVSVV KVDEIQKQGE NNPVKALQGR VPGMNITADG N PSGSATVR IRGIGTLNNN DPLYIIDGVP TKAGMHELNG NDIESIQVLK DAASASIYGS RAANGVIIIT TKQGKKGQIK IN FDASVSA SMYQSKMNVL NTEQYGRAMW QAYVNDGENP NGNALGYAYN WGYNADGNPV LYGMTLSKYL DSKNTMPVAD TDW FDEITR TGVIQQYNLS VSNGSEKGSS FFSLGYYKNL GVIKDTDFDR FSARMNSDYK LIDDILTIGQ HFTLNRTSEV QAPG GIIET ALDIPSAIPV YASDGSWGGP VGGWPDRRNP RAVLEYNKDN RYTYWRMFGD AYVNLTPFKG FNLRSTFGLD YANKQ ARYF TYPYQEGTQT NNGKSAVEAK QEHWTKWMWN AIATYQLEVG KHRGDVMIGM ELNREDDSHF SGYKEDFSIL TPDYMW PDA GSGTAQAYGA GEGYSLVSFF GKMNYSYADR YLLSLTLRRD GSSRFGKNHR YATFPSVSLG WRITQENFMK ELTWLDD LK LRASWGQTGN QEISNLARYT IYAPNYGTTD SFGGQSYGTA YDITGSNGGG VLPSGFKRNQ IGNDNIKWET TTQTNVGI D FSLFKQSLYG SLEYYYKKAT DILTEMAGVG VLGEGGSRWI NSGAMKNQGF EFNLGYRNKT AFGLTYDLNG NISTYRNEI LELPETVAAN GKFGGNGVKS VVGHTYGAQV GYIADGIFKS QDEVDNHATQ EGAAVGRIRY RDIDHNGVID ERDQNWIYDP TPSFSYGLN IYLEYKNFDL TMFWQGVQGV DIISDVKKKS DFWSASNVGF LNKGTRLLNA WSPTNPNSDI PALTRSDTNN E QRVSTYFV ENGSFLKLRN IQLGYTVPAV ISKKMRMDRL RFYCSAQNLL TIKSKNFTGE DPENPNFSYP IPVNITFGLN IG F |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.02 mg/mL | ||||||||||||
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Buffer | pH: 7.5 Component:
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 63.84 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |