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Yorodumi- EMDB-3096: cryo-ET subtomogram averaging of BG505 SOSIP.664 in complex with ... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-3096 | |||||||||
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Title | cryo-ET subtomogram averaging of BG505 SOSIP.664 in complex with sCD4, 17b, and 8ANC195 | |||||||||
Map data | subtomogram-averaging of 1745 sub-volumes. | |||||||||
Sample |
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Function / homology | Function and homology information helper T cell enhancement of adaptive immune response / interleukin-16 binding / interleukin-16 receptor activity / maintenance of protein location in cell / T cell selection / MHC class II protein binding / cellular response to granulocyte macrophage colony-stimulating factor stimulus / interleukin-15-mediated signaling pathway / positive regulation of monocyte differentiation / Nef Mediated CD4 Down-regulation ...helper T cell enhancement of adaptive immune response / interleukin-16 binding / interleukin-16 receptor activity / maintenance of protein location in cell / T cell selection / MHC class II protein binding / cellular response to granulocyte macrophage colony-stimulating factor stimulus / interleukin-15-mediated signaling pathway / positive regulation of monocyte differentiation / Nef Mediated CD4 Down-regulation / Alpha-defensins / positive regulation of kinase activity / regulation of T cell activation / T cell receptor complex / extracellular matrix structural constituent / Other interleukin signaling / enzyme-linked receptor protein signaling pathway / Dectin-2 family / Translocation of ZAP-70 to Immunological synapse / Phosphorylation of CD3 and TCR zeta chains / regulation of calcium ion transport / macrophage differentiation / Generation of second messenger molecules / T cell differentiation / PD-1 signaling / positive regulation of protein kinase activity / Binding and entry of HIV virion / coreceptor activity / positive regulation of calcium-mediated signaling / positive regulation of plasma membrane raft polarization / positive regulation of receptor clustering / protein tyrosine kinase binding / positive regulation of establishment of T cell polarity / positive regulation of interleukin-2 production / virus-mediated perturbation of host defense response / T cell activation / host cell endosome membrane / Vpu mediated degradation of CD4 / calcium-mediated signaling / clathrin-coated endocytic vesicle membrane / cell surface receptor protein tyrosine kinase signaling pathway / positive regulation of T cell activation / positive regulation of peptidyl-tyrosine phosphorylation / transmembrane signaling receptor activity / Cargo recognition for clathrin-mediated endocytosis / Downstream TCR signaling / virus receptor activity / signaling receptor activity / MHC class II protein complex binding / Clathrin-mediated endocytosis / clathrin-dependent endocytosis of virus by host cell / positive regulation of canonical NF-kappaB signal transduction / defense response to Gram-negative bacterium / positive regulation of MAPK cascade / adaptive immune response / positive regulation of viral entry into host cell / cell surface receptor signaling pathway / positive regulation of ERK1 and ERK2 cascade / viral protein processing / early endosome / cell adhesion / immune response / positive regulation of protein phosphorylation / membrane raft / fusion of virus membrane with host plasma membrane / external side of plasma membrane / endoplasmic reticulum lumen / fusion of virus membrane with host endosome membrane / viral envelope / lipid binding / endoplasmic reticulum membrane / virion attachment to host cell / protein kinase binding / host cell plasma membrane / virion membrane / structural molecule activity / positive regulation of DNA-templated transcription / enzyme binding / signal transduction / protein homodimerization activity / zinc ion binding / membrane / identical protein binding / plasma membrane Similarity search - Function | |||||||||
Biological species | Human immunodeficiency virus 1 / Homo sapiens (human) | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 23.0 Å | |||||||||
Authors | Scharf L / Wang H / Gao H / Chen S / McDowall A / Bjorkman P | |||||||||
Citation | Journal: Cell / Year: 2015 Title: Broadly Neutralizing Antibody 8ANC195 Recognizes Closed and Open States of HIV-1 Env. Authors: Louise Scharf / Haoqing Wang / Han Gao / Songye Chen / Alasdair W McDowall / Pamela J Bjorkman / Abstract: The HIV-1 envelope (Env) spike contains limited epitopes for broadly neutralizing antibodies (bNAbs); thus, most neutralizing antibodies are strain specific. The 8ANC195 epitope, defined by crystal ...The HIV-1 envelope (Env) spike contains limited epitopes for broadly neutralizing antibodies (bNAbs); thus, most neutralizing antibodies are strain specific. The 8ANC195 epitope, defined by crystal and electron microscopy (EM) structures of bNAb 8ANC195 complexed with monomeric gp120 and trimeric Env, respectively, spans the gp120 and gp41 Env subunits. To investigate 8ANC195's gp41 epitope at higher resolution, we solved a 3.58 Å crystal structure of 8ANC195 complexed with fully glycosylated Env trimer, revealing 8ANC195 insertion into a glycan shield gap to contact gp120 and gp41 glycans and protein residues. To determine whether 8ANC195 recognizes the CD4-bound open Env conformation that leads to co-receptor binding and fusion, one of several known conformations of virion-associated Env, we solved EM structures of an Env/CD4/CD4-induced antibody/8ANC195 complex. 8ANC195 binding partially closed the CD4-bound trimer, confirming structural plasticity of Env by revealing a previously unseen conformation. 8ANC195's ability to bind different Env conformations suggests advantages for potential therapeutic applications. | |||||||||
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_3096.map.gz | 351.6 KB | EMDB map data format | |
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Header (meta data) | emd-3096-v30.xml emd-3096.xml | 13.9 KB 13.9 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_3096_fsc.xml | 2.2 KB | Display | FSC data file |
Images | emd-3096.jpg | 1.1 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-3096 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-3096 | HTTPS FTP |
-Related structure data
Related structure data | 5a8hMC 3086C 5a7xC 5cjxC M: atomic model generated by this map C: citing same article (ref.) |
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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_3096.map.gz / Format: CCP4 / Size: 478.5 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | subtomogram-averaging of 1745 sub-volumes. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 5.2 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : soluble HIV-1 Env trimer BG505 SOSIP.664 in complex with soluble ...
Entire | Name: soluble HIV-1 Env trimer BG505 SOSIP.664 in complex with soluble CD4s (D1-D2), broadly neutralizing antibody 17b Fabs, and broadly neutralizing antibody 8ANC195 variant G32K5 Fabs |
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Components |
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-Supramolecule #1000: soluble HIV-1 Env trimer BG505 SOSIP.664 in complex with soluble ...
Supramolecule | Name: soluble HIV-1 Env trimer BG505 SOSIP.664 in complex with soluble CD4s (D1-D2), broadly neutralizing antibody 17b Fabs, and broadly neutralizing antibody 8ANC195 variant G32K5 Fabs type: sample / ID: 1000 Oligomeric state: 3 Fabs of 17b, 3 sCD4, and 3 Fabs of 8ANC195 G52K5 bind to Env trimer BG505 SOSIP Number unique components: 4 |
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Molecular weight | Theoretical: 500 KDa |
-Macromolecule #1: Soluble HIV-1 Env trimer BG505 SOSIP.664
Macromolecule | Name: Soluble HIV-1 Env trimer BG505 SOSIP.664 / type: protein_or_peptide / ID: 1 / Name.synonym: BG505 SOSIP / Number of copies: 1 / Oligomeric state: trimer / Recombinant expression: Yes |
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Source (natural) | Organism: Human immunodeficiency virus 1 / synonym: HIV-1 |
Molecular weight | Theoretical: 420 KDa |
Recombinant expression | Organism: Homo sapiens (human) / Recombinant cell: HEK293-6E / Recombinant plasmid: pTT5 |
-Macromolecule #2: Fab of broadly neutralizing antibody 17b
Macromolecule | Name: Fab of broadly neutralizing antibody 17b / type: protein_or_peptide / ID: 2 / Name.synonym: 17b / Number of copies: 3 / Oligomeric state: monomer / Recombinant expression: Yes |
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Source (natural) | Organism: Homo sapiens (human) / synonym: Human |
Molecular weight | Theoretical: 50 KDa |
Recombinant expression | Organism: Homo sapiens (human) / Recombinant cell: HEK293-6E / Recombinant plasmid: pTT5 |
-Macromolecule #3: soluble CD4 (D1-D2 domains)
Macromolecule | Name: soluble CD4 (D1-D2 domains) / type: protein_or_peptide / ID: 3 / Name.synonym: sCD4 / Number of copies: 3 / Oligomeric state: monomer / Recombinant expression: Yes |
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Source (natural) | Organism: Homo sapiens (human) / synonym: Human |
Molecular weight | Theoretical: 20 KDa |
Recombinant expression | Organism: Homo sapiens (human) / Recombinant cell: HEK293-6E / Recombinant plasmid: pTT5 |
-Macromolecule #4: Fab of broadly neutralizing antibody 8ANC195 variant G52K5
Macromolecule | Name: Fab of broadly neutralizing antibody 8ANC195 variant G52K5 type: protein_or_peptide / ID: 4 / Name.synonym: 8ANC195 G52K5 / Number of copies: 3 / Oligomeric state: monomer / Recombinant expression: Yes |
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Source (natural) | Organism: Homo sapiens (human) / synonym: Human |
Molecular weight | Theoretical: 50 KDa |
Recombinant expression | Organism: Homo sapiens (human) / Recombinant cell: HEK293-6E / Recombinant plasmid: pTT5 |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | particle |
-Sample preparation
Concentration | 0.06 mg/mL |
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Buffer | pH: 8 / Details: 20mM Tris, 50mM NaCl |
Grid | Details: Quantifoil R2/2 NH2 copper finder mesh with extra thick carbon grid |
Vitrification | Cryogen name: ETHANE-PROPANE MIXTURE / Chamber humidity: 95 % / Chamber temperature: 70 K / Instrument: FEI VITROBOT MARK IV / Method: blot for 2.5 sec, 6 blotting force |
-Electron microscopy
Microscope | FEI POLARA 300 |
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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: 2.2 mm / Nominal magnification: 41000 |
Specialist optics | Energy filter - Name: Gatan energy filter |
Sample stage | Specimen holder model: GATAN HELIUM / Tilt series - Axis1 - Min angle: -60 ° / Tilt series - Axis1 - Max angle: 60 ° |
Date | Jan 25, 2015 |
Image recording | Category: CCD / Film or detector model: GATAN K2 (4k x 4k) / Number real images: 25 / Average electron dose: 120 e/Å2 |
Experimental equipment | Model: Tecnai Polara / Image courtesy: FEI Company |
-Image processing
-Atomic model buiding 1
Initial model | PDB ID: Chain - #0 - Chain ID: G / Chain - #1 - Chain ID: C / Chain - #2 - Chain ID: L / Chain - #3 - Chain ID: H |
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Software | Name: Chimera |
Refinement | Space: REAL / Protocol: RIGID BODY FIT |
Output model | PDB-5a8h: |