<?xml version="1.0" encoding="UTF-8"?>
<emd xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:noNamespaceSchemaLocation="https://ftp.ebi.ac.uk/pub/databases/em_ebi/emdb_related/emdb-schemas/emdb_schemas/v3/v3_0_9_3/emdb.xsd" version="3.0.9.3" emdb_id="EMD-8564">
    <admin>
        <current_status>
            <date>2024-10-23</date>
            <code>REL</code>
            <processing_site>RCSB</processing_site>
        </current_status>
        <sites>
            <deposition>RCSB</deposition>
            <last_processing>RCSB</last_processing>
        </sites>
        <key_dates>
            <deposition>2017-01-19</deposition>
            <header_release>2017-02-15</header_release>
            <map_release>2017-05-31</map_release>
            <update>2024-10-23</update>
        </key_dates>
        <grant_support>
            <grant_reference>
                <funding_body>National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)</funding_body>
                <code>P01 AI-089618</code>
                <country>United States</country>
            </grant_reference>
        </grant_support>
        <title>CryoEM structure of an influenza virus receptor-binding site antibody-antigen interface - Class 4</title>
        <authors_list>
            <author>Liu Y</author>
            <author>Pan J</author>
        </authors_list>
        <keywords>viral glycoprotein, hemagglutinin, antibody fragment, VIRAL PROTEIN-IMMUNE SYSTEM complex</keywords>
    </admin>
    <crossreferences>
        <citation_list>
            <primary_citation>
                <journal_citation published="true">
                    <author order="1">Liu Y</author>
                    <author order="2">Pan J</author>
                    <author order="3">Jenni S</author>
                    <author order="4">Raymond DD</author>
                    <author order="5">Caradonna T</author>
                    <author order="6">Do KT</author>
                    <author order="7">Schmidt AG</author>
                    <author order="8">Harrison SC</author>
                    <author order="9">Grigorieff N</author>
                    <title>CryoEM Structure of an Influenza Virus Receptor-Binding Site Antibody-Antigen Interface.</title>
                    <journal_abbreviation>J. Mol. Biol.</journal_abbreviation>
                    <country>UK</country>
                    <volume>429</volume>
                    <first_page>1829</first_page>
                    <last_page>1839</last_page>
                    <year>2017</year>
                    <external_references type="PUBMED">28506635</external_references>
                    <external_references type="DOI">doi:10.1016/j.jmb.2017.05.011</external_references>
                    <external_references type="ISSN">1089-8638</external_references>
                    <external_references type="CSD">0070</external_references>
                    <external_references type="ASTM">JMOBAK</external_references>
                </journal_citation>
            </primary_citation>
        </citation_list>
        <emdb_list>
            <emdb_reference>
                <emdb_id>EMD-8561</emdb_id>
                <relationship>
                    <other>other EM volume</other>
                </relationship>
            </emdb_reference>
            <emdb_reference>
                <emdb_id>EMD-8562</emdb_id>
                <relationship>
                    <other>other EM volume</other>
                </relationship>
            </emdb_reference>
            <emdb_reference>
                <emdb_id>EMD-8563</emdb_id>
                <relationship>
                    <other>other EM volume</other>
                </relationship>
            </emdb_reference>
        </emdb_list>
        <pdb_list>
            <pdb_reference>
                <pdb_id>5uk2</pdb_id>
                <relationship>
                    <in_frame>FULLOVERLAP</in_frame>
                </relationship>
            </pdb_reference>
        </pdb_list>
    </crossreferences>
    <sample>
        <name>Influenza-virus hemagglutinin H1 complexed with K1915 single-chain variable-domain fragment</name>
        <supramolecule_list>
            <complex_supramolecule supramolecule_id="1">
                <name>Influenza-virus hemagglutinin H1 complexed with K1915 single-chain variable-domain fragment</name>
                <parent>0</parent>
                <macromolecule_list>
                    <macromolecule>
                        <macromolecule_id>1</macromolecule_id>
                    </macromolecule>
                    <macromolecule>
                        <macromolecule_id>2</macromolecule_id>
                    </macromolecule>
                    <macromolecule>
                        <macromolecule_id>3</macromolecule_id>
                    </macromolecule>
                </macromolecule_list>
                <details>The complex consists of three hemagglutinin head domains bound to three single-chain variable-domain fragments.</details>
                <natural_source database="NCBI">
                    <organism ncbi="464623">Influenza A virus (A/Solomon Islands/3/2006(H1N1))</organism>
                </natural_source>
                <molecular_weight>
                    <theoretical units="MDa">0.32</theoretical>
                </molecular_weight>
            </complex_supramolecule>
        </supramolecule_list>
        <macromolecule_list>
            <protein_or_peptide macromolecule_id="1">
                <name>Hemagglutinin HA1</name>
                <natural_source database="NCBI">
                    <organism ncbi="464623">Influenza A virus (A/Solomon Islands/3/2006(H1N1))</organism>
                    <strain>A/Solomon Islands/3/2006(H1N1)</strain>
                </natural_source>
                <molecular_weight>
                    <theoretical units="MDa">0.03602434399999999</theoretical>
                </molecular_weight>
                <number_of_copies>3</number_of_copies>
                <recombinant_expression database="NCBI">
                    <recombinant_organism ncbi="7111">Trichoplusia ni</recombinant_organism>
                </recombinant_expression>
                <enantiomer>LEVO</enantiomer>
                <sequence>
                    <string>EDTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCLLKGIAPLQLGNCSVAGWILGNPECELLISRESWSYI
VEKPNPENGTCYPGHFADYEELREQLSSVSSFERFEIFPKESSWPNHTTTGVSASCSHNGESSFYKNLLWLTGKNGLYPN
LSKSYANNKEKEVLVLWGVHHPPNIGDQRALYHTENAYVSVVSSHYSRKFTPEIAKRPKVRDREGRINYYWTLLEPGDTI
IFEANGNLIAPRYAFALSRGFGSGIINSNAPMDECDAKCQTPQGAINSSLPFQNVHPVTIGECPKYVRSAKLRMVTGLRN
IPS</string>
                    <external_references type="UNIPROTKB">A7UPX0</external_references>
                </sequence>
            </protein_or_peptide>
            <protein_or_peptide macromolecule_id="2">
                <name>Hemagglutinin HA2</name>
                <natural_source database="NCBI">
                    <organism ncbi="464623">Influenza A virus</organism>
                    <strain>A/Solomon Islands/3/2006(H1N1)</strain>
                </natural_source>
                <molecular_weight>
                    <theoretical units="MDa">0.019841041</theoretical>
                </molecular_weight>
                <number_of_copies>3</number_of_copies>
                <recombinant_expression database="NCBI">
                    <recombinant_organism ncbi="7111">Trichoplusia ni</recombinant_organism>
                </recombinant_expression>
                <enantiomer>LEVO</enantiomer>
                <sequence>
                    <string>GLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADQKSTQNAINGITNKVNSVIEKMNTQFTAVGKEFNKLERRMENL
NKKVDDGFIDIWTYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNNAKEIGNGCFEFYHKCNDECMESVKNGTYDYP
KYSEESKLNREKI</string>
                    <external_references type="UNIPROTKB">A7UPX0</external_references>
                </sequence>
            </protein_or_peptide>
            <protein_or_peptide macromolecule_id="3">
                <name>scFv</name>
                <natural_source database="NCBI">
                    <organism ncbi="9606">Homo sapiens</organism>
                </natural_source>
                <molecular_weight>
                    <theoretical units="MDa">0.027629384999999996</theoretical>
                </molecular_weight>
                <number_of_copies>3</number_of_copies>
                <recombinant_expression database="NCBI">
                    <recombinant_organism ncbi="9606">Homo sapiens</recombinant_organism>
                </recombinant_expression>
                <enantiomer>LEVO</enantiomer>
                <sequence>
                    <string>EIVLTQSPGTLSLSPGERATLSCRASQSISGYYLTWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLE
PEDFAVYYCQQYGISPVITFGGGTNVEIKGGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFNIYDMH
WVRQAPGKGLEWVSGLTTGGDTSYSGSVRGRFSISRENAKNSLYLQMNNLRAGDTAAYFCVRGVREVGATGGDPFYYAMA
VWGQGTTVTVSSASGSSGSGHHHHHH</string>
                </sequence>
            </protein_or_peptide>
            <ligand macromolecule_id="6">
                <name>2-acetamido-2-deoxy-beta-D-glucopyranose</name>
                <molecular_weight>
                    <theoretical units="MDa">0.000221208</theoretical>
                </molecular_weight>
                <number_of_copies>18</number_of_copies>
                <formula>NAG</formula>
            </ligand>
        </macromolecule_list>
    </sample>
    <structure_determination_list>
        <structure_determination structure_determination_id="1">
            <method>singleParticle</method>
            <aggregation_state>particle</aggregation_state>
            <specimen_preparation_list>
                <single_particle_preparation preparation_id="1">
                    <concentration units="mg/mL">0.1</concentration>
                    <buffer>
                        <ph>8.0</ph>
                        <details>Beta-octylglucoside was added to a final concentration of 0.07% w/v to induce more variable particle orientations.</details>
                    </buffer>
                    <grid>
                        <model>Quantifoil R1.2/1.3</model>
                        <material>COPPER</material>
                        <mesh>200</mesh>
                        <support_film film_type_id="1">
                            <film_material>CARBON</film_material>
                            <film_topology>HOLEY ARRAY</film_topology>
                        </support_film>
                        <pretreatment>
                            <type>GLOW DISCHARGE</type>
                            <atmosphere>AIR</atmosphere>
                        </pretreatment>
                    </grid>
                    <vitrification>
                        <cryogen_name>ETHANE</cryogen_name>
                        <chamber_humidity units="percentage">90</chamber_humidity>
                        <chamber_temperature units="K">298</chamber_temperature>
                        <instrument>FEI VITROBOT MARK I</instrument>
                    </vitrification>
                </single_particle_preparation>
            </specimen_preparation_list>
            <microscopy_list>
                <single_particle_microscopy microscopy_id="1">
                    <microscope>FEI TITAN KRIOS</microscope>
                    <illumination_mode>FLOOD BEAM</illumination_mode>
                    <imaging_mode>BRIGHT FIELD</imaging_mode>
                    <electron_source>FIELD EMISSION GUN</electron_source>
                    <acceleration_voltage units="kV">300</acceleration_voltage>
                    <nominal_cs units="mm">2.7</nominal_cs>
                    <calibrated_defocus_min units="µm">1.0</calibrated_defocus_min>
                    <calibrated_defocus_max units="µm">9.0</calibrated_defocus_max>
                    <nominal_magnification>18000.0</nominal_magnification>
                    <calibrated_magnification>30444.0</calibrated_magnification>
                    <specimen_holder_model>FEI TITAN KRIOS AUTOGRID HOLDER</specimen_holder_model>
                    <cooling_holder_cryogen>NITROGEN</cooling_holder_cryogen>
                    <temperature>
                        <temperature_min units="K">70.0</temperature_min>
                        <temperature_max units="K">70.0</temperature_max>
                    </temperature>
                    <alignment_procedure>
                        <basic/>
                    </alignment_procedure>
                    <image_recording_list>
                        <image_recording image_recording_id="1">
                            <film_or_detector_model>GATAN K2 SUMMIT (4k x 4k)</film_or_detector_model>
                            <detector_mode>SUPER-RESOLUTION</detector_mode>
                            <digitization_details>
                                <dimensions>
                                    <width units="pixel">7676</width>
                                    <height units="pixel">7420</height>
                                </dimensions>
                                <frames_per_image>1-38</frames_per_image>
                            </digitization_details>
                            <number_grids_imaged>1</number_grids_imaged>
                            <number_real_images>10281</number_real_images>
                            <average_exposure_time units="s">13.0</average_exposure_time>
                            <average_electron_dose_per_image units="e/Å^2">40.0</average_electron_dose_per_image>
                            <details>The exposure rate was 8 electrons/physical pixel/second.</details>
                        </image_recording>
                    </image_recording_list>
                </single_particle_microscopy>
            </microscopy_list>
            <singleparticle_processing image_processing_id="1">
                <image_recording_id>1</image_recording_id>
                <details>The particle images were normalized to have constant variance and zero average. Movies were processed using Unblur.</details>
                <particle_selection>
                    <number_selected>252130</number_selected>
                </particle_selection>
                <startup_model type_of_model="OTHER">
                    <details>A starting map was determined using rosettes formed from Ni-NTA nanogold and His-tagged particles.</details>
                </startup_model>
                <final_reconstruction>
                    <number_classes_used>3</number_classes_used>
                    <algorithm>FOURIER SPACE</algorithm>
                    <resolution units="Å" res_type="BY AUTHOR">4.8</resolution>
                    <resolution_method>FSC 0.143 CUT-OFF</resolution_method>
                    <software_list>
                        <software>
                            <name>FREALIGN</name>
                            <version>9.11</version>
                        </software>
                    </software_list>
                    <details>Refinement and classification were limited to 10 Angstrom resolution.</details>
                    <number_images_used>142314</number_images_used>
                </final_reconstruction>
                <initial_angle_assignment>
                    <type>PROJECTION MATCHING</type>
                    <projection_matching_processing>
                        <angular_sampling units="degrees">7.5</angular_sampling>
                    </projection_matching_processing>
                    <software_list>
                        <software>
                            <name>IMAGIC</name>
                        </software>
                    </software_list>
                </initial_angle_assignment>
                <final_angle_assignment>
                    <type>PROJECTION MATCHING</type>
                    <projection_matching_processing/>
                    <software_list>
                        <software>
                            <name>FREALIGN</name>
                            <version>9.11</version>
                        </software>
                    </software_list>
                </final_angle_assignment>
                <final_three_d_classification>
                    <number_classes>4</number_classes>
                    <average_number_members_per_class>35579.0</average_number_members_per_class>
                    <software_list>
                        <software>
                            <name>FREALIGN</name>
                            <version>9.11</version>
                        </software>
                    </software_list>
                    <details>Three out of the four classes showed high-resolution features.</details>
                </final_three_d_classification>
            </singleparticle_processing>
        </structure_determination>
    </structure_determination_list>
    <map format="CCP4" size_kbytes="6913">
        <file>emd_8564.map.gz</file>
        <symmetry>
            <space_group>1</space_group>
        </symmetry>
        <data_type>IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)</data_type>
        <dimensions>
            <col>120</col>
            <row>120</row>
            <sec>120</sec>
        </dimensions>
        <origin>
            <col>-60</col>
            <row>-60</row>
            <sec>-60</sec>
        </origin>
        <spacing>
            <x>120</x>
            <y>120</y>
            <z>120</z>
        </spacing>
        <cell>
            <a units="Å">196.8</a>
            <b units="Å">196.8</b>
            <c units="Å">196.8</c>
            <alpha units="deg">90.0</alpha>
            <beta units="deg">90.0</beta>
            <gamma units="deg">90.0</gamma>
        </cell>
        <axis_order>
            <fast>X</fast>
            <medium>Y</medium>
            <slow>Z</slow>
        </axis_order>
        <statistics>
            <minimum>-3.741372</minimum>
            <maximum>8.258274999999999</maximum>
            <average>0.071280874</average>
            <std>0.45758796</std>
        </statistics>
        <pixel_spacing>
            <x units="Å">1.64</x>
            <y units="Å">1.64</y>
            <z units="Å">1.64</z>
        </pixel_spacing>
        <contour_list>
            <contour primary="true">
                <level>2.52</level>
                <source>EMDB</source>
            </contour>
        </contour_list>
        <label>::::EMDATABANK.org::::EMD-8564::::</label>
        <annotation_details>influenza virus receptor-binding site antibody-antigen interface - Class 4</annotation_details>
    </map>
    <interpretation>
        <modelling_list>
            <modelling>
                <refinement_protocol>OTHER</refinement_protocol>
                <details>Domains were initially placed manually using Chimera. The HA trimer was from PDB ID 5UGY.  Fab was initially obtained using MODELLER, with PDB ID 4K8R as template. The structure was refined using PHENIX.</details>
                <target_criteria>Correlation coefficient</target_criteria>
                <refinement_space>REAL</refinement_space>
            </modelling>
        </modelling_list>
    </interpretation>
</emd>
