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Yorodumi- EMDB-45754: The cryo-EM structure of BamACDE complex from Neisseria gonorrhoeae -
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Open data
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Basic information
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| Title | The cryo-EM structure of BamACDE complex from Neisseria gonorrhoeae | ||||||||||||||||||
Map data | cryo-EM map of BamACDE from Neisseria gonorrhoeae- final map | ||||||||||||||||||
Sample |
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Keywords | beta barrel assembly machinery / BAM complex / outer membrane protein / Neisseria / Neisseria gonnorhoeae / protein folding / beta barrel membrane protein / MEMBRANE PROTEIN | ||||||||||||||||||
| Function / homology | Function and homology informationestablishment of competence for transformation / Bam protein complex / Gram-negative-bacterium-type cell outer membrane assembly / protein insertion into membrane / cell outer membrane / protein-macromolecule adaptor activity / metal ion binding Similarity search - Function | ||||||||||||||||||
| Biological species | Neisseria gonorrhoeae (bacteria) / Neisseria gonorrhoeae FA 1090 (bacteria) | ||||||||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 4.0 Å | ||||||||||||||||||
Authors | Billings EM / Noinaj N | ||||||||||||||||||
| Funding support | United States, 5 items
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Citation | Journal: Structure / Year: 2025Title: Structural insights into outer membrane protein biogenesis in pathogenic Neisseria. Authors: Evan Billings / Zixing Fan / Moloud Aflaki Sooreshjani / James C Gumbart / Nicholas Noinaj / ![]() Abstract: N. gonorrhoeae (Ngo) causes the sexually transmitted infection gonorrhea with ∼106 million infections worldwide annually. Ngo infections can result in an increased risk of acquiring HIV, ...N. gonorrhoeae (Ngo) causes the sexually transmitted infection gonorrhea with ∼106 million infections worldwide annually. Ngo infections can result in an increased risk of acquiring HIV, infertility, and blindness. To combat Ngo infections, we report the cryoelectron microscopy (cryo-EM) structure of the Ngo β-barrel assembly machinery (NgBAM), which is responsible for the biogenesis of β-barrel outer membrane proteins (OMPs). NgBAM was observed in an inward-open state; however, the polypeptide transport-associated (POTRA) domains more closely match those found in the outward-open state in E. coli β-barrel assembly machinery (BAM). The barrel seam of NgBamA consists of partial pairing of strand β1 with β16; no outward-open state of NgBAM was observed. Molecular dynamics (MD) simulations reveal unique overall dynamics and interplay between the POTRA domains of NgBamA and NgBamD. We propose that in Ngo, initial recognition occurs in the inward-open state where the last strand of the OMP partially pairs with β1 of NgBamA and must compete off β16. | ||||||||||||||||||
| History |
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Structure visualization
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_45754.map.gz | 59.7 MB | EMDB map data format | |
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| Header (meta data) | emd-45754-v30.xml emd-45754.xml | 30.8 KB 30.8 KB | Display Display | EMDB header |
| Images | emd_45754.png | 38.6 KB | ||
| Filedesc metadata | emd-45754.cif.gz | 8.3 KB | ||
| Others | emd_45754_half_map_1.map.gz emd_45754_half_map_2.map.gz | 59.5 MB 59.5 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-45754 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-45754 | HTTPS FTP |
-Validation report
| Summary document | emd_45754_validation.pdf.gz | 828.2 KB | Display | EMDB validaton report |
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| Full document | emd_45754_full_validation.pdf.gz | 827.7 KB | Display | |
| Data in XML | emd_45754_validation.xml.gz | 12.1 KB | Display | |
| Data in CIF | emd_45754_validation.cif.gz | 14.3 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-45754 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-45754 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9cmwMC ![]() 9cn0C ![]() 9cn1C M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_45754.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | cryo-EM map of BamACDE from Neisseria gonorrhoeae- final map | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.069 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Half map: cryo-EM map of BamACDE from Neisseria gonorrhoeae- half map A
| File | emd_45754_half_map_1.map | ||||||||||||
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| Annotation | cryo-EM map of BamACDE from Neisseria gonorrhoeae- half map A | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: cryo-EM map of BamACDE from Neisseria gonorrhoeae- half map B
| File | emd_45754_half_map_2.map | ||||||||||||
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| Annotation | cryo-EM map of BamACDE from Neisseria gonorrhoeae- half map B | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
-Entire : NgBamACDE
| Entire | Name: NgBamACDE |
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| Components |
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-Supramolecule #1: NgBamACDE
| Supramolecule | Name: NgBamACDE / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all Details: beta barrel assembly machnery complex from Neisseria gonorrhoeae |
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| Source (natural) | Organism: Neisseria gonorrhoeae (bacteria) / Strain: FA1090 |
| Molecular weight | Theoretical: 183 kDa/nm |
-Macromolecule #1: Outer membrane protein assembly factor BamA
| Macromolecule | Name: Outer membrane protein assembly factor BamA / type: protein_or_peptide / ID: 1 Details: The endogenous signal sequence in the NgBamA gene was replaced by the pelB signal sequence, which is cleaved in the mature protein Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Neisseria gonorrhoeae FA 1090 (bacteria) / Strain: FA1090 |
| Molecular weight | Theoretical: 88.025969 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MGKYLLPTAA AGLLLLAAQP AMDFTIQDIR VEGLQRTEPS TVFNYLPVKV GDTYNDTHGS AIIKSLYATG FFDDVRVETA DGQLLLTVI ERPTIGSLNI TGAKMLQNDA IKKNLESFGL AQSQYFNQAT LNQAVAGLKE EYLGRGKLNI QITPKVTKLA R NRVDIDIT ...String: MGKYLLPTAA AGLLLLAAQP AMDFTIQDIR VEGLQRTEPS TVFNYLPVKV GDTYNDTHGS AIIKSLYATG FFDDVRVETA DGQLLLTVI ERPTIGSLNI TGAKMLQNDA IKKNLESFGL AQSQYFNQAT LNQAVAGLKE EYLGRGKLNI QITPKVTKLA R NRVDIDIT IDEGKSAKIT DIEFEGNQVY SDRKLMRQMS LTEGGIWTWL TRSDRFDRQK FAQDMEKVTD FYQNNGYFDF RI LDTDIQT NEDKTRQTIK ITVHEGGRFR WGKVSIEGDT NEVPKAELEK LLTMKPGKWY ERQQMTAVLG EIQNRMGSAG YAY SEISVQ PLPNAGTKTV DFVLHIEPGR KIYVNEIHIT GNNKTRDEVV RRELRQMESA PYDTSKLQRS KERVELLGYF DNVQ FDAVP LAGTPDKVDL NMSLTERSTG SLDLSAGWVQ DTGLVMSAGV SQDNLFGTGK SAALRASRSK TTLNGSLSFT DPYFT ADGV SLGYDIYGKA FDPRKASTSV KQYKTTTAGG GVRMGIPVTE YDRVNFGLAA EHLTVNTYNK APKRYADFIR KYGKTD GAD GSFKGLLYKG TVGWGRNKTD SASWPTRGYL TGVNAEIALP GSKLQYYSAT HNQTWFFPLS KTFTLMLGGE VGIAGGY GR TKEIPFFENF YGGGLGSVRG YESGTLGPKV YDEYGEKISY GGNKKANVSA ELLFPMPGAK DARTVRLSLF ADAGSVWD G RTYTAAENGN NKSVYSENAH KSTFTNELRY SAGGAVTWLS PLGPMKFSYA YPLKKKPEDE IQRFQFQLGT TF UniProtKB: Outer membrane protein assembly factor BamA |
-Macromolecule #2: Outer membrane protein assembly factor bamC
| Macromolecule | Name: Outer membrane protein assembly factor bamC / type: protein_or_peptide / ID: 2 Details: The endogenous lipidation signal sequence was replaced with the lipidation signal sequence from E. coli BamB Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Neisseria gonorrhoeae FA 1090 (bacteria) / Strain: FA1090 |
| Molecular weight | Theoretical: 41.946664 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MGQLRKLLLP GLLSVTLLSG CGMGSGSKTE QPKLDYQSRS HRLIKLEVPP DLNNPDQGNL YRLPAGSGAV RASDLEKRRT PAVQQPADA EVLKSVKGVR LERDGSQRWL VVDGKSPAEI WPLLKAFWQE NGFDIESEEP AIGQMETEWA ENRAKIPQDS L RRLFDTVG ...String: MGQLRKLLLP GLLSVTLLSG CGMGSGSKTE QPKLDYQSRS HRLIKLEVPP DLNNPDQGNL YRLPAGSGAV RASDLEKRRT PAVQQPADA EVLKSVKGVR LERDGSQRWL VVDGKSPAEI WPLLKAFWQE NGFDIESEEP AIGQMETEWA ENRAKIPQDS L RRLFDTVG LGGIYSTGER DKFIVRIEQG KNGVSDIFFA HKAMKEVYGD KNKDTTMWQP SASDPNLEAA FLTRFMQYLG VD GRQAENA LAKKPTLPAA NEMARIEGKS LIVFGDYGRN WRRTGLALDR IGLTVVGQNT ERHAFLVQKA PNESNAVTEQ KPG LFKRLL GKGKAEKPAE QPELIVYAEP VADGSRIVLL NKDGSAYAGK DASALLGKLH SELR UniProtKB: Lipoprotein |
-Macromolecule #3: Outer membrane protein assembly factor BamD
| Macromolecule | Name: Outer membrane protein assembly factor BamD / type: protein_or_peptide / ID: 3 Details: The endogenous lipidation signal sequence was replaced with the lipidation signal sequence from E. coli BamB Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Neisseria gonorrhoeae FA 1090 (bacteria) / Strain: FA1090 |
| Molecular weight | Theoretical: 31.535693 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MQLRKLLLPG LLSVTLLSGC AMGATQGTAD KDAQITQDWS VEKLYAEAQD ELNSSNYTRA VKLYEILESR FPTSRHARQS QLDTAYAYY KDDEKDKALA AIERFRRLHP QHPNMDYALY LRGLVLFNED QSFLNKLASQ DWSDRDPKAN REAYQAFAEL V QRFPNSKY ...String: MQLRKLLLPG LLSVTLLSGC AMGATQGTAD KDAQITQDWS VEKLYAEAQD ELNSSNYTRA VKLYEILESR FPTSRHARQS QLDTAYAYY KDDEKDKALA AIERFRRLHP QHPNMDYALY LRGLVLFNED QSFLNKLASQ DWSDRDPKAN REAYQAFAEL V QRFPNSKY AADATARMVK LVDALGGNEM SVARYYMKRG AYIAAANRAK KIIGSYQNTR YVEESLAILE LAYKKLDKPQ LA ADTRRVL ETNFPKSPFL THAWQPDDMP WWRYWH UniProtKB: Outer membrane protein assembly factor BamD |
-Macromolecule #4: Outer membrane protein assembly factor BamE
| Macromolecule | Name: Outer membrane protein assembly factor BamE / type: protein_or_peptide / ID: 4 Details: The endogenous lipidation signal sequence was replaced with the lipidation sequence sequence from E. coli BamB. the C-terminus contains a 10x histadine tag. Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: Neisseria gonorrhoeae FA 1090 (bacteria) / Strain: FA1090 |
| Molecular weight | Theoretical: 15.915229 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MQLRKLLLPG LLSVTLLSGC AMDSVERVSL FPSYKLKIIQ GNELEPRAVA ALRPGMTKDQ VLLLLGSPIL RDAFHTDRWD YTFNTSRNG IIKERSNLTV YFENGVLVRT EGDALQNAAE ALRAKQNADK QHHHHHHHHH H UniProtKB: Outer membrane protein assembly factor BamE |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 4.2 mg/mL | ||||||||||||||||||
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| Buffer | pH: 7.5 Component:
Details: 1xPBS pH 7.5, with 0.01% LMNG | ||||||||||||||||||
| Grid | Model: Quantifoil R3.5/1 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec. / Pretreatment - Atmosphere: AIR | ||||||||||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: Blot force of 2, blot time of 15 seconds.. | ||||||||||||||||||
| Details | Sample used directly after purification over a Superdex200 size exclusion chromatography column. |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Specialist optics | Energy filter - Slit width: 20 eV |
| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Digitization - Dimensions - Width: 11520 pixel / Digitization - Dimensions - Height: 8184 pixel / Number grids imaged: 1 / Number real images: 10825 / Average exposure time: 0.03 sec. / Average electron dose: 41.87 e/Å2 Details: Data was collected at the National Center for Cryo-EM Access and Training, using one of the Titan Krios (Elizabeth). |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 9.700000000000001 µm / Nominal defocus min: 2.23 µm / Nominal magnification: 81000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Initial model |
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| Refinement | Space: REAL / Protocol: RIGID BODY FIT | ||||||||
| Output model | ![]() PDB-9cmw: |
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About Yorodumi



Keywords
Neisseria gonorrhoeae (bacteria)
Authors
United States, 5 items
Citation









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FIELD EMISSION GUN



