+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-43249 | |||||||||
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Title | TehA from Haemophilus influenzae purified in OG | |||||||||
Map data | ||||||||||
Sample |
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Keywords | ANION CHANNEL / ALPHA HELICAL INTEGRAL MEMBRANE PROTEIN / MEMBRANE PROTEIN | |||||||||
Function / homology | Function and homology information monoatomic cation efflux transmembrane transporter activity / response to tellurium ion / response to antibiotic / identical protein binding / plasma membrane Similarity search - Function | |||||||||
Biological species | Haemophilus influenzae (bacteria) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | |||||||||
Authors | Catalano C / Senko S / Tran NL / Lucier KW / Farwell AC / Silva MS / Dip PV / Poweleit N / Scapin G | |||||||||
Funding support | 1 items
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Citation | Journal: Int J Mol Sci / Year: 2024 Title: High-Resolution Cryo-Electron Microscopy Structure Determination of Tellurite-Resistance Protein A via 200 kV Transmission Electron Microscopy. Authors: Nhi L Tran / Skerdi Senko / Kyle W Lucier / Ashlyn C Farwell / Sabrina M Silva / Phat V Dip / Nicole Poweleit / Giovanna Scapin / Claudio Catalano / Abstract: Membrane proteins constitute about 20% of the human proteome and play crucial roles in cellular functions. However, a complete understanding of their structure and function is limited by their ...Membrane proteins constitute about 20% of the human proteome and play crucial roles in cellular functions. However, a complete understanding of their structure and function is limited by their hydrophobic nature, which poses significant challenges in purification and stabilization. Detergents, essential in the isolation process, risk destabilizing or altering the proteins' native conformations, thus affecting stability and functionality. This study leverages single-particle cryo-electron microscopy to elucidate the structural nuances of membrane proteins, focusing on the SLAC1 bacterial homolog from (TehA) purified with diverse detergents, including n-dodecyl β-D-maltopyranoside (DDM), glycodiosgenin (GDN), β-D-octyl-glucoside (OG), and lauryl maltose neopentyl glycol (LMNG). This research not only contributes to the understanding of membrane protein structures but also addresses detergent effects on protein purification. By showcasing that the overall structural integrity of the channel is preserved, our study underscores the intricate interplay between proteins and detergents, offering insightful implications for drug design and membrane biology. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_43249.map.gz | 306.7 MB | EMDB map data format | |
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Header (meta data) | emd-43249-v30.xml emd-43249.xml | 16.5 KB 16.5 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_43249_fsc.xml | 14.6 KB | Display | FSC data file |
Images | emd_43249.png | 35.7 KB | ||
Masks | emd_43249_msk_1.map | 325 MB | Mask map | |
Filedesc metadata | emd-43249.cif.gz | 5.9 KB | ||
Others | emd_43249_half_map_1.map.gz emd_43249_half_map_2.map.gz | 301.7 MB 301.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-43249 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-43249 | HTTPS FTP |
-Validation report
Summary document | emd_43249_validation.pdf.gz | 1 MB | Display | EMDB validaton report |
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Full document | emd_43249_full_validation.pdf.gz | 1 MB | Display | |
Data in XML | emd_43249_validation.xml.gz | 23.2 KB | Display | |
Data in CIF | emd_43249_validation.cif.gz | 29.6 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43249 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43249 | HTTPS FTP |
-Related structure data
Related structure data | 8vi5MC 8vi2C 8vi3C 8vi4C M: atomic model generated by this map 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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-Map
File | Download / File: emd_43249.map.gz / Format: CCP4 / Size: 325 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.566 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_43249_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_43249_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_43249_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : TELLURITE RESISTANCE PROTEIN TEHA
Entire | Name: TELLURITE RESISTANCE PROTEIN TEHA |
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Components |
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-Supramolecule #1: TELLURITE RESISTANCE PROTEIN TEHA
Supramolecule | Name: TELLURITE RESISTANCE PROTEIN TEHA / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Haemophilus influenzae (bacteria) |
Molecular weight | Theoretical: 108 kDa/nm |
-Macromolecule #1: Tellurite resistance protein TehA homolog
Macromolecule | Name: Tellurite resistance protein TehA homolog / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO |
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Source (natural) | Organism: Haemophilus influenzae (bacteria) |
Molecular weight | Theoretical: 35.252547 KDa |
Recombinant expression | Organism: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria) |
Sequence | String: MNITKPFPLP TGYFGIPLGL AALSLAWFHL ENLFPAARMV SDVLGIVASA VWILFILMYA YKLRYYFEEV RAEYHSPVRF SFIALIPIT TMLVGDILYR WNPLIAEVLI WIGTIGQLLF STLRVSELWQ GGVFEQKSTH PSFYLPAVAA NFTSASSLAL L GYHDLGYL ...String: MNITKPFPLP TGYFGIPLGL AALSLAWFHL ENLFPAARMV SDVLGIVASA VWILFILMYA YKLRYYFEEV RAEYHSPVRF SFIALIPIT TMLVGDILYR WNPLIAEVLI WIGTIGQLLF STLRVSELWQ GGVFEQKSTH PSFYLPAVAA NFTSASSLAL L GYHDLGYL FFGAGMIAWI IFEPVLLQHL RISSLEPQFR ATMGIVLAPA FVCVSAYLSI NHGEVDTLAK ILWGYGFLQL FF LLRLFPW IVEKGLNIGL WAFSFGLASM ANSATAFYHG NVLQGVSIFA FVFSNVMIGL LVLMTIYKLT KGQFFLK UniProtKB: Tellurite resistance protein TehA homolog |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 3 mg/mL |
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Buffer | pH: 8 |
Grid | Model: UltrAuFoil R1.2/1.3 / Material: GOLD / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec. |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | TFS GLACIOS |
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Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Number real images: 8354 / Average electron dose: 36.43 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.5 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 240000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |