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Yorodumi- EMDB-17787: 4.0 angstrom map of outward-facing MFS transporter MHAS2168, a ho... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-17787 | ||||||||||||
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Title | 4.0 angstrom map of outward-facing MFS transporter MHAS2168, a homologue of M. tuberculosis Rv1410, in complex with a megabody | ||||||||||||
Map data | Membrane transporter MHAS2168, a homologue of M. tuberculosis Rv1410, in complex with megabody H2 consisting of nanobody H2 grafted to HopQ domain | ||||||||||||
Sample |
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Keywords | Major Facilitator Superfamily transporter / megabody / outward-open conformation / triacylglyceride extraction / MEMBRANE PROTEIN | ||||||||||||
Function / homology | Function and homology information | ||||||||||||
Biological species | Mycolicibacterium hassiacum DSM 44199 (bacteria) / Vicugna pacos (alpaca) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.99 Å | ||||||||||||
Authors | Remm S / Gonda I / Seeger MA | ||||||||||||
Funding support | Switzerland, European Union, 3 items
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Citation | Journal: Nat Commun / Year: 2023 Title: Structural basis for triacylglyceride extraction from mycobacterial inner membrane by MFS transporter Rv1410. Authors: Sille Remm / Dario De Vecchis / Jendrik Schöppe / Cedric A J Hutter / Imre Gonda / Michael Hohl / Simon Newstead / Lars V Schäfer / Markus A Seeger / Abstract: Mycobacterium tuberculosis is protected from antibiotic therapy by a multi-layered hydrophobic cell envelope. Major facilitator superfamily (MFS) transporter Rv1410 and the periplasmic lipoprotein ...Mycobacterium tuberculosis is protected from antibiotic therapy by a multi-layered hydrophobic cell envelope. Major facilitator superfamily (MFS) transporter Rv1410 and the periplasmic lipoprotein LprG are involved in transport of triacylglycerides (TAGs) that seal the mycomembrane. Here, we report a 2.7 Å structure of a mycobacterial Rv1410 homologue, which adopts an outward-facing conformation and exhibits unusual transmembrane helix 11 and 12 extensions that protrude ~20 Å into the periplasm. A small, very hydrophobic cavity suitable for lipid transport is constricted by a functionally important ion-lock likely involved in proton coupling. Combining mutational analyses and MD simulations, we propose that TAGs are extracted from the core of the inner membrane into the central cavity via lateral clefts present in the inward-facing conformation. The functional role of the periplasmic helix extensions is to channel the extracted TAG into the lipid binding pocket of LprG. | ||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_17787.map.gz | 327.8 MB | EMDB map data format | |
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Header (meta data) | emd-17787-v30.xml emd-17787.xml | 19.9 KB 19.9 KB | Display Display | EMDB header |
Images | emd_17787.png | 60.2 KB | ||
Masks | emd_17787_msk_1.map | 347.6 MB | Mask map | |
Filedesc metadata | emd-17787.cif.gz | 6.4 KB | ||
Others | emd_17787_half_map_1.map.gz emd_17787_half_map_2.map.gz | 323.1 MB 323.1 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-17787 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-17787 | HTTPS FTP |
-Related structure data
Related structure data | 8pnlC C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_17787.map.gz / Format: CCP4 / Size: 347.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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Annotation | Membrane transporter MHAS2168, a homologue of M. tuberculosis Rv1410, in complex with megabody H2 consisting of nanobody H2 grafted to HopQ domain | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.65 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_17787_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_17787_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_17787_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : MFS transporter MHAS2168 in outward-open conformation in complex ...
Entire | Name: MFS transporter MHAS2168 in outward-open conformation in complex with Megabody_H2 |
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Components |
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-Supramolecule #1: MFS transporter MHAS2168 in outward-open conformation in complex ...
Supramolecule | Name: MFS transporter MHAS2168 in outward-open conformation in complex with Megabody_H2 type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Mycolicibacterium hassiacum DSM 44199 (bacteria) |
-Macromolecule #1: MFS transporter MHAS2168, a homologue of M. tuberculosis Rv1410c,...
Macromolecule | Name: MFS transporter MHAS2168, a homologue of M. tuberculosis Rv1410c, in outward-open conformation type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Mycolicibacterium hassiacum DSM 44199 (bacteria) |
Recombinant expression | Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) |
Sequence | String: MSAFPQTPNR LIRPRRTSRG IAISAGGLAV LLGALDTYVV VSIVTDIMRD VGIAVNQIQR VTPIITGYLL GYIAAMPLLG RASDRFGRKL LIQISLAGFA LGSVITALAT NLDVLVAGRV IQGAASGALL PVTLALAADL WATHKRAAVL GGVGAAQELG AVLGPIYGIF ...String: MSAFPQTPNR LIRPRRTSRG IAISAGGLAV LLGALDTYVV VSIVTDIMRD VGIAVNQIQR VTPIITGYLL GYIAAMPLLG RASDRFGRKL LIQISLAGFA LGSVITALAT NLDVLVAGRV IQGAASGALL PVTLALAADL WATHKRAAVL GGVGAAQELG AVLGPIYGIF VVWLFHHWQA VFWVNVPLAL IAMVLIHISL PPRVRTEEPQ RVDVTGGLLL ALALGLATIG LYNAEPDGKQ VLPEYGPPLI IGAVIAAVAF LVWERFARTR LLDPAGVRFR PFLIALLVSL VTGGALMVTL VNVELFGQGV LGLDQDEAVF LLARFLIALP VGALLGGWIA TRVGDRAVTA VGLLIAAGGF YLIAQWPADV LESRHDLGFV SLPTLDTDLA IAGFGLGLVI APLTSAALRV VPAAQHGIAS AAVVVARMIG MLIGIAALSA WGLYRFNQYL KEQLAALPPA PADFPGGQMA GQMMRLRTAT VQAYVLQYGE IFAITAGLCV FGAVLGLFIA GRREHAEESA DAVDGVSNAR DRAPSAALEV LFQ UniProtKB: Putative triacylglyceride transporter |
-Macromolecule #2: Megabody_H2, derived from grafting alpaca-produced Nanobody_H2 on...
Macromolecule | Name: Megabody_H2, derived from grafting alpaca-produced Nanobody_H2 onto the HopQ adhesin domain of Helicobacter pylori type: protein_or_peptide / ID: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: Vicugna pacos (alpaca) |
Recombinant expression | Organism: Escherichia coli MC1061 (bacteria) |
Sequence | String: QVQLQESGGG LVQTKTTTSV IDTTNDAQNL LTQAQTIVNT LKDYCPILIA KSSSSNGGTN NANTPSWQTA GGGKNSCATF GAEFSAASDM INNAQKIVQE TQQLSANQPK NITQPHNLNL NSPSSLTALA QKMLKNAQSQ AEILKLANQV ESDFNKLSSG HLKDYIGKCD ...String: QVQLQESGGG LVQTKTTTSV IDTTNDAQNL LTQAQTIVNT LKDYCPILIA KSSSSNGGTN NANTPSWQTA GGGKNSCATF GAEFSAASDM INNAQKIVQE TQQLSANQPK NITQPHNLNL NSPSSLTALA QKMLKNAQSQ AEILKLANQV ESDFNKLSSG HLKDYIGKCD ASAISSANMT MQNQKNNWGN GCAGVEETQS LLKTSAADFN NQTPQINQAQ NLANTLIQEL GNNPFRGGGK LSDTYEQLSR LLTNDNGTNS KTSAQAINQA VNNLNERAKT LAGGTTNSPA YQATLLALRS VLGLWNSMGY AVICGGYTKS PGENNQKDFH YTDENGNGTT INCGGSTNSN GTHSYNGTNT LKADKNVSLS IEQYEKIHEA YQILSKALKQ AGLAPLNSKG EKLEAHVTTS KYGSPGGSLR LSCADAGSIF NKFPMAWYRQ APGKERELVA RISSGGSTNY ADFVKGRFTI SRDNAKSTLY LQMNSLKPED TAMYYCARII NSASNIAYWG QGTRVTVSAA LEVLFQ |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 9.4 mg/mL | ||||||||||||
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Buffer | pH: 7.4 Component:
Details: 20 mM Tris/HCl pH 7.4, 150 mM NaCl, 0.03% (w/v) beta-DDM | ||||||||||||
Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 45 sec. | ||||||||||||
Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 283.15 K / Instrument: LEICA PLUNGER | ||||||||||||
Details | This sample was monodisperse |
-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 / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 130000 |
Sample stage | Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number grids imaged: 3 / Number real images: 11713 / Average electron dose: 66.54 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Particle selection | Number selected: 4759395 Details: Particles extracted after template picking and manual inspection of particle picks |
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Startup model | Type of model: NONE |
Initial angle assignment | Type: ANGULAR RECONSTITUTION / Software - Name: cryoSPARC (ver. v3.2) |
Final 3D classification | Number classes: 2 / Avg.num./class: 273034 / Software - Name: cryoSPARC (ver. v3.2) / Software - details: Heterogeneous refinement Details: The best class had 402229 particles and worst class had 143839 particles. |
Final angle assignment | Type: ANGULAR RECONSTITUTION / Software - Name: cryoSPARC (ver. v3.2) |
Final reconstruction | Number classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 3.99 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. v3.2) / Software - details: Non-uniform refinement / Number images used: 402229 |