[English] 日本語
Yorodumi- EMDB-23152: Cryo-electron microscopy reconstruction of antibody DH898.1 Fab-d... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-23152 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Cryo-electron microscopy reconstruction of antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of HIV Env CH848 10.17 SOSIP trimer | |||||||||
Map data | Final sharpened, locally filtered, map of antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of HIV Env CH848 10.17 SOSIP trimer | |||||||||
Sample |
| |||||||||
Function / homology | Function and homology information positive regulation of plasma membrane raft polarization / positive regulation of receptor clustering / positive regulation of establishment of T cell polarity / host cell endosome membrane / clathrin-dependent endocytosis of virus by host cell / viral protein processing / fusion of virus membrane with host plasma membrane / virus-mediated perturbation of host defense response / fusion of virus membrane with host endosome membrane / viral envelope ...positive regulation of plasma membrane raft polarization / positive regulation of receptor clustering / positive regulation of establishment of T cell polarity / host cell endosome membrane / clathrin-dependent endocytosis of virus by host cell / viral protein processing / fusion of virus membrane with host plasma membrane / virus-mediated perturbation of host defense response / fusion of virus membrane with host endosome membrane / viral envelope / virion attachment to host cell / host cell plasma membrane / structural molecule activity / virion membrane / plasma membrane Similarity search - Function | |||||||||
Biological species | Human immunodeficiency virus 1 / Macaca mulatta (Rhesus monkey) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 6.1 Å | |||||||||
Authors | Edwards RJ / Acharya P | |||||||||
Funding support | United States, 2 items
| |||||||||
Citation | Journal: Cell / Year: 2021 Title: Fab-dimerized glycan-reactive antibodies are a structural category of natural antibodies. Authors: Wilton B Williams / R Ryan Meyerhoff / R J Edwards / Hui Li / Kartik Manne / Nathan I Nicely / Rory Henderson / Ye Zhou / Katarzyna Janowska / Katayoun Mansouri / Sophie Gobeil / Tyler ...Authors: Wilton B Williams / R Ryan Meyerhoff / R J Edwards / Hui Li / Kartik Manne / Nathan I Nicely / Rory Henderson / Ye Zhou / Katarzyna Janowska / Katayoun Mansouri / Sophie Gobeil / Tyler Evangelous / Bhavna Hora / Madison Berry / A Yousef Abuahmad / Jordan Sprenz / Margaret Deyton / Victoria Stalls / Megan Kopp / Allen L Hsu / Mario J Borgnia / Guillaume B E Stewart-Jones / Matthew S Lee / Naomi Bronkema / M Anthony Moody / Kevin Wiehe / Todd Bradley / S Munir Alam / Robert J Parks / Andrew Foulger / Thomas Oguin / Gregory D Sempowski / Mattia Bonsignori / Celia C LaBranche / David C Montefiori / Michael Seaman / Sampa Santra / John Perfect / Joseph R Francica / Geoffrey M Lynn / Baptiste Aussedat / William E Walkowicz / Richard Laga / Garnett Kelsoe / Kevin O Saunders / Daniela Fera / Peter D Kwong / Robert A Seder / Alberto Bartesaghi / George M Shaw / Priyamvada Acharya / Barton F Haynes / Abstract: Natural antibodies (Abs) can target host glycans on the surface of pathogens. We studied the evolution of glycan-reactive B cells of rhesus macaques and humans using glycosylated HIV-1 envelope (Env) ...Natural antibodies (Abs) can target host glycans on the surface of pathogens. We studied the evolution of glycan-reactive B cells of rhesus macaques and humans using glycosylated HIV-1 envelope (Env) as a model antigen. 2G12 is a broadly neutralizing Ab (bnAb) that targets a conserved glycan patch on Env of geographically diverse HIV-1 strains using a unique heavy-chain (V) domain-swapped architecture that results in fragment antigen-binding (Fab) dimerization. Here, we describe HIV-1 Env Fab-dimerized glycan (FDG)-reactive bnAbs without V-swapped domains from simian-human immunodeficiency virus (SHIV)-infected macaques. FDG Abs also recognized cell-surface glycans on diverse pathogens, including yeast and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike. FDG precursors were expanded by glycan-bearing immunogens in macaques and were abundant in HIV-1-naive humans. Moreover, FDG precursors were predominately mutated IgMIgDCD27, thus suggesting that they originated from a pool of antigen-experienced IgM or marginal zone B cells. #1: Journal: bioRxiv / Year: 2020 Title: Fab-dimerized glycan-reactive antibodies neutralize HIV and are prevalent in humans and rhesus macaques Authors: Williams WB / Meyerhoff RR / Edwards RJ / Li H / Nicely NI / Henderson R / Zhou Y / Janowska K / Mansouri K / Manne K / Stalls V / Hsu AL / Borgnia MJ / Stewart-Jones G / Lee MS / Bronkema N ...Authors: Williams WB / Meyerhoff RR / Edwards RJ / Li H / Nicely NI / Henderson R / Zhou Y / Janowska K / Mansouri K / Manne K / Stalls V / Hsu AL / Borgnia MJ / Stewart-Jones G / Lee MS / Bronkema N / Perfect J / Moody MA / Wiehe K / Bradley T / Kepler TB / Alam SM / Foulger A / Bonsignori M / LaBranche CC / Montefiori DC / Seaman M / Santra S / Francica JR / Lynn GM / Aussedat B / Walkowicz WE / Laga R / Kelso G / Saunders KO / Fera D / Kwong PD / Seder RA / Bartesaghi A / Shaw GM / Acharya P / Haynes BF | |||||||||
History |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_23152.map.gz | 6.4 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-23152-v30.xml emd-23152.xml | 33.1 KB 33.1 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_23152_fsc.xml | 11.2 KB | Display | FSC data file |
Images | emd_23152.png | 158.7 KB | ||
Masks | emd_23152_msk_1.map emd_23152_msk_2.map | 125 MB 125 MB | Mask map | |
Others | emd_23152_additional_1.map.gz emd_23152_additional_2.map.gz emd_23152_half_map_1.map.gz emd_23152_half_map_2.map.gz | 61.4 MB 6.1 MB 116 MB 116 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-23152 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-23152 | HTTPS FTP |
-Validation report
Summary document | emd_23152_validation.pdf.gz | 532.3 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_23152_full_validation.pdf.gz | 531.9 KB | Display | |
Data in XML | emd_23152_validation.xml.gz | 18.9 KB | Display | |
Data in CIF | emd_23152_validation.cif.gz | 24.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23152 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23152 | HTTPS FTP |
-Related structure data
Related structure data | 6vtuC 6xrjC 7l02C 7l06C 7l09C 7l6mC 7l6oC 7lu9C 7luaC C: citing same article (ref.) |
---|---|
Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_23152.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Final sharpened, locally filtered, map of antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of HIV Env CH848 10.17 SOSIP trimer | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.0665 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Mask #1
File | emd_23152_msk_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Mask #2
File | emd_23152_msk_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Additional map: Unmodified final map of antibody DH898.1 Fab-dimer bound...
File | emd_23152_additional_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Unmodified final map of antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of HIV Env CH848 10.17 SOSIP trimer | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Additional map: Local resolution of final map of antibody DH898.1...
File | emd_23152_additional_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Local resolution of final map of antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of HIV Env CH848 10.17 SOSIP trimer | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: Half-map A of antibody DH898.1 Fab-dimer bound to...
File | emd_23152_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Half-map A of antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of HIV Env CH848 10.17 SOSIP trimer | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: Half-map B of antibody DH898.1 Fab-dimer bound to...
File | emd_23152_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Half-map B of antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of HIV Env CH848 10.17 SOSIP trimer | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Sample components
-Entire : Antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of ...
Entire | Name: Antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of HIV Env CH848.3.D0949.10.17chim.6R.DS.SOSIP.664 trimer |
---|---|
Components |
|
-Supramolecule #1: Antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of ...
Supramolecule | Name: Antibody DH898.1 Fab-dimer bound to glycans 332, 392, and 396 of HIV Env CH848.3.D0949.10.17chim.6R.DS.SOSIP.664 trimer type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
---|---|
Molecular weight | Theoretical: 94 KDa |
-Supramolecule #2: HIV Env CH848.3.D0949.10.17chim.6R.DS.SOSIP.664 trimer
Supramolecule | Name: HIV Env CH848.3.D0949.10.17chim.6R.DS.SOSIP.664 trimer type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
---|---|
Source (natural) | Organism: Human immunodeficiency virus 1 / Strain: CH848 10.17 |
Recombinant expression | Organism: Homo sapiens (human) / Recombinant cell: 293F |
-Supramolecule #3: Antibody DH898.1 Fragment antigen binding (Fab) dimer
Supramolecule | Name: Antibody DH898.1 Fragment antigen binding (Fab) dimer / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2-#3 / Details: Composed of two heavy chains and two light chains |
---|---|
Source (natural) | Organism: Macaca mulatta (Rhesus monkey) |
Recombinant expression | Organism: Homo sapiens (human) / Recombinant cell: 293F |
-Macromolecule #1: HIV Env CH848.3.D0949.10.17chim.6R.DS.SOSIP.664
Macromolecule | Name: HIV Env CH848.3.D0949.10.17chim.6R.DS.SOSIP.664 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Human immunodeficiency virus 1 / Strain: CH848.3.D0949.10.17 |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: AENLWVTVYY GVPVWKEAKT TLFCASDARA YEKEVHNVWA THACVPTDPS PQELVLGNVT ENFNMWKNDM VDQMHEDIIS LWDQSLKPCV KLTPLCVTLI CSNATVKNGT VEEMKNCSFN TTTEIRDKEK KEYALFYKPD IVPLSETNNT SEYRLINCNT SACTQACPKV ...String: AENLWVTVYY GVPVWKEAKT TLFCASDARA YEKEVHNVWA THACVPTDPS PQELVLGNVT ENFNMWKNDM VDQMHEDIIS LWDQSLKPCV KLTPLCVTLI CSNATVKNGT VEEMKNCSFN TTTEIRDKEK KEYALFYKPD IVPLSETNNT SEYRLINCNT SACTQACPKV TFEPIPIHYC APAGYAILKC NDETFNGTGP CSNVSTVQCT HGIRPVVSTQ LLLNGSLAEK EIVIRSENLT NNAKIIIVHL HTPVEIVCTR PNNNTRKSVR IGPGQTFYAT GDIIGDIKQA HCNISEEKWN DTLQKVGIEL QKHFPNKTIK YNQSAGGDME ITTHSFNCGG EFFYCNTSNL FNGTYNGTYI STNSSANSTS TITLQCRIKQ IINMWQGVGR CMYAPPIAGN ITCRSNITGL LLTRDGGTNS NETETFRPAG GDMRDNWRSE LYKYKVVKIE PLGVAPTRCK RRVVGRRRRR RAVGIGAVFL GFLGAAGSTM GAASMTLTVQ ARNLLSGIVQ QQSNLLRAPE AQQHLLKLTV WGIKQLQARV LAVERYLRDQ QLLGIWGCSG KLICCTNVPW NSSWSNRNLS EIWDNMTWLQ WDKEISNYTQ IIYGLLEESQ NQQEKNEQDL LALD |
-Macromolecule #2: DH898.1 Fab heavy chain
Macromolecule | Name: DH898.1 Fab heavy chain / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Macaca mulatta (Rhesus monkey) |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: QDLLLQSGAE VREPGASVTV SCQASNYTFP DYYIHWVRLV PGQGLEWLGE MKPKVGVTNV SKKIRDRLFM TADTSTDTAY MVLSALTPGD TAIYYCTRLE PDFLSGWAHW GKGVLVTVSP ASTKGPSVFP LAPSSRSTSE STAALGCLVK DYFPEPVTVS WNSGSLTSGV ...String: QDLLLQSGAE VREPGASVTV SCQASNYTFP DYYIHWVRLV PGQGLEWLGE MKPKVGVTNV SKKIRDRLFM TADTSTDTAY MVLSALTPGD TAIYYCTRLE PDFLSGWAHW GKGVLVTVSP ASTKGPSVFP LAPSSRSTSE STAALGCLVK DYFPEPVTVS WNSGSLTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTQT YVCNVNHKPS NTKVDKRVE |
-Macromolecule #3: DH898.1 Fab light chain
Macromolecule | Name: DH898.1 Fab light chain / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Macaca mulatta (Rhesus monkey) |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: EIVMTQTPLS LSVTPGEPAS LSCRSSASLL HGNGNTYLHW YLRKAGQSPQ LLIFGGSKRV PGISDRFIGS GAGTNFTLKI SSVEADDVGF YYCAQGVAFP WTFGQGTKVE IKRAVAAPSV FIFPPSEDQV KSGTVSVVCL LNNFYPREAS VKWKVDGVLK TGNSQESVTE ...String: EIVMTQTPLS LSVTPGEPAS LSCRSSASLL HGNGNTYLHW YLRKAGQSPQ LLIFGGSKRV PGISDRFIGS GAGTNFTLKI SSVEADDVGF YYCAQGVAFP WTFGQGTKVE IKRAVAAPSV FIFPPSEDQV KSGTVSVVCL LNNFYPREAS VKWKVDGVLK TGNSQESVTE QDSKDNTYSL SSTLTLSNTD YQSHNVYACE VTHQGLSSPV TKSFNRGE |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 1.5 mg/mL | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Buffer | pH: 7.5 Component:
| |||||||||
Grid | Model: Quantifoil R1.2/1.3 / Support film - Material: CARBON / Support film - topology: HOLEY ARRAY / Support film - Film thickness: 12.0 nm | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 295 K / Instrument: LEICA EM GP Details: 2.5 microliters of sample applied to grid and incubated for 30 seconds in chamber, blotted for 2.5 seconds, and plunge frozen in liquid ethane. | |||||||||
Details | For the DH898.1-bound complex, purified HIV-1 Env was mixed with 6-fold molar excess of recombinantly expressed and purified DH898.1 Fab to obtain a final Env concentration of 1.5 mg/ml, and incubated for 2 hours at room temperature. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number real images: 3230 / Average exposure time: 4.0 sec. / Average electron dose: 60.0 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: 3.5 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 22500 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |