Importance of the Side Chain at Position 296 of Antibody Fc in Interactions with FcγRIIIa and Other Fcγ Receptors
Antibody-dependent cellular cytotoxicity (ADCC) is an important effector function determining the clinical efficacy of therapeutic antibodies. Core fucose removal from N-glycans on the Fc portion of immunoglobulin G (IgG) improves the binding affinity for Fcγ receptor IIIa (FcγRIIIa) and dramaticall...
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| description | Antibody-dependent cellular cytotoxicity (ADCC) is an important effector function determining the clinical efficacy of therapeutic antibodies. Core fucose removal from N-glycans on the Fc portion of immunoglobulin G (IgG) improves the binding affinity for Fcγ receptor IIIa (FcγRIIIa) and dramatically enhances ADCC. Our previous structural analyses revealed that Tyr-296 of IgG1-Fc plays a critical role in the interaction with FcγRIIIa, particularly in the enhanced FcγRIIIa binding of nonfucosylated IgG1. However, the importance of the Tyr-296 residue in the antibody in the interaction with various Fcγ receptors has not yet been elucidated. To further clarify the biological importance of this residue, we established comprehensive Tyr-296 mutants as fucosylated and nonfucosylated anti-CD20 IgG1s rituximab variants and examined their binding to recombinant soluble human Fcγ receptors: shFcγRI, shFcγRIIa, shFcγRIIIa, and shFcγRIIIb. Some of the mutations affected the binding of antibody to not only shFcγRIIIa but also shFcγRIIa and shFcγRIIIb, suggesting that the Tyr-296 residue in the antibody was also involved in interactions with FcγRIIa and FcγRIIIb. For FcγRIIIa binding, almost all Tyr-296 variants showed lower binding affinities than the wild-type antibody, irrespective of their core fucosylation, particularly in Y296K and Y296P. Notably, only the Y296W mutant showed improved binding to FcγRIIIa. The 3.00 Å-resolution crystal structure of the nonfucosylated Y296W mutant in complex with shFcγRIIIa harboring two N-glycans revealed that the Tyr-to-Trp substitution increased the number of potential contact atoms in the complex, thus improving the binding of the antibody to shFcγRIIIa. The nonfucosylated Y296W mutant retained high ADCC activity, relative to the nonfucosylated wild-type IgG1, and showed greater binding affinity for FcγRIIa. Our data may improve our understanding of the biological importance of human IgG1-Fc Tyr-296 in interactions with various Fcγ receptors, and have applications in the modulation of the IgG1-Fc function of therapeutic antibodies. |
| doi_str_mv | 10.1371/journal.pone.0140120 |
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Core fucose removal from N-glycans on the Fc portion of immunoglobulin G (IgG) improves the binding affinity for Fcγ receptor IIIa (FcγRIIIa) and dramatically enhances ADCC. Our previous structural analyses revealed that Tyr-296 of IgG1-Fc plays a critical role in the interaction with FcγRIIIa, particularly in the enhanced FcγRIIIa binding of nonfucosylated IgG1. However, the importance of the Tyr-296 residue in the antibody in the interaction with various Fcγ receptors has not yet been elucidated. To further clarify the biological importance of this residue, we established comprehensive Tyr-296 mutants as fucosylated and nonfucosylated anti-CD20 IgG1s rituximab variants and examined their binding to recombinant soluble human Fcγ receptors: shFcγRI, shFcγRIIa, shFcγRIIIa, and shFcγRIIIb. Some of the mutations affected the binding of antibody to not only shFcγRIIIa but also shFcγRIIa and shFcγRIIIb, suggesting that the Tyr-296 residue in the antibody was also involved in interactions with FcγRIIa and FcγRIIIb. For FcγRIIIa binding, almost all Tyr-296 variants showed lower binding affinities than the wild-type antibody, irrespective of their core fucosylation, particularly in Y296K and Y296P. Notably, only the Y296W mutant showed improved binding to FcγRIIIa. The 3.00 Å-resolution crystal structure of the nonfucosylated Y296W mutant in complex with shFcγRIIIa harboring two N-glycans revealed that the Tyr-to-Trp substitution increased the number of potential contact atoms in the complex, thus improving the binding of the antibody to shFcγRIIIa. The nonfucosylated Y296W mutant retained high ADCC activity, relative to the nonfucosylated wild-type IgG1, and showed greater binding affinity for FcγRIIa. Our data may improve our understanding of the biological importance of human IgG1-Fc Tyr-296 in interactions with various Fcγ receptors, and have applications in the modulation of the IgG1-Fc function of therapeutic antibodies.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0140120</identifier><identifier>PMID: 26444434</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Affinity ; Animals ; Antibodies ; Antibody-Dependent Cell Cytotoxicity ; Antigens, CD20 - immunology ; Atomic properties ; Binding ; CD20 antigen ; Cell Line ; CHO Cells ; Contact potentials ; Cricetulus ; Crystal structure ; Crystallography, X-Ray ; Cytotoxicity ; Epidermal growth factor ; Fucose ; Fucose - chemistry ; Fucose - genetics ; Fucose - immunology ; Humans ; Immunoglobulin Fc Fragments - chemistry ; Immunoglobulin Fc Fragments - genetics ; Immunoglobulin Fc Fragments - immunology ; Immunoglobulin G ; Immunoglobulins ; Immunology ; Leukemia ; Lymphoma ; Lymphomas ; Models, Molecular ; Monoclonal antibodies ; Mutants ; Mutation ; N-glycans ; Pharmaceutical sciences ; Point Mutation ; Polysaccharides ; R&D ; Receptor mechanisms ; Receptors ; Receptors, IgG - immunology ; Research & development ; Research parks ; Rituximab ; Rituximab - chemistry ; Rituximab - genetics ; Rituximab - immunology ; Targeted cancer therapy ; Toxicity</subject><ispartof>PloS one, 2015-10, Vol.10 (10), p.e0140120-e0140120</ispartof><rights>2015 Isoda et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2015 Isoda et al 2015 Isoda et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-cec17a510bd8e686d1abc4a5f916c4d7dd8d4af5867498580703a58620a598053</citedby><cites>FETCH-LOGICAL-c526t-cec17a510bd8e686d1abc4a5f916c4d7dd8d4af5867498580703a58620a598053</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4596520/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4596520/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26444434$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Barchi, Joseph J</contributor><creatorcontrib>Isoda, Yuya</creatorcontrib><creatorcontrib>Yagi, Hirokazu</creatorcontrib><creatorcontrib>Satoh, Tadashi</creatorcontrib><creatorcontrib>Shibata-Koyama, Mami</creatorcontrib><creatorcontrib>Masuda, Kazuhiro</creatorcontrib><creatorcontrib>Satoh, Mitsuo</creatorcontrib><creatorcontrib>Kato, Koichi</creatorcontrib><creatorcontrib>Iida, Shigeru</creatorcontrib><title>Importance of the Side Chain at Position 296 of Antibody Fc in Interactions with FcγRIIIa and Other Fcγ Receptors</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Antibody-dependent cellular cytotoxicity (ADCC) is an important effector function determining the clinical efficacy of therapeutic antibodies. Core fucose removal from N-glycans on the Fc portion of immunoglobulin G (IgG) improves the binding affinity for Fcγ receptor IIIa (FcγRIIIa) and dramatically enhances ADCC. Our previous structural analyses revealed that Tyr-296 of IgG1-Fc plays a critical role in the interaction with FcγRIIIa, particularly in the enhanced FcγRIIIa binding of nonfucosylated IgG1. However, the importance of the Tyr-296 residue in the antibody in the interaction with various Fcγ receptors has not yet been elucidated. To further clarify the biological importance of this residue, we established comprehensive Tyr-296 mutants as fucosylated and nonfucosylated anti-CD20 IgG1s rituximab variants and examined their binding to recombinant soluble human Fcγ receptors: shFcγRI, shFcγRIIa, shFcγRIIIa, and shFcγRIIIb. Some of the mutations affected the binding of antibody to not only shFcγRIIIa but also shFcγRIIa and shFcγRIIIb, suggesting that the Tyr-296 residue in the antibody was also involved in interactions with FcγRIIa and FcγRIIIb. For FcγRIIIa binding, almost all Tyr-296 variants showed lower binding affinities than the wild-type antibody, irrespective of their core fucosylation, particularly in Y296K and Y296P. Notably, only the Y296W mutant showed improved binding to FcγRIIIa. The 3.00 Å-resolution crystal structure of the nonfucosylated Y296W mutant in complex with shFcγRIIIa harboring two N-glycans revealed that the Tyr-to-Trp substitution increased the number of potential contact atoms in the complex, thus improving the binding of the antibody to shFcγRIIIa. The nonfucosylated Y296W mutant retained high ADCC activity, relative to the nonfucosylated wild-type IgG1, and showed greater binding affinity for FcγRIIa. Our data may improve our understanding of the biological importance of human IgG1-Fc Tyr-296 in interactions with various Fcγ receptors, and have applications in the modulation of the IgG1-Fc function of therapeutic antibodies.</description><subject>Affinity</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Antibody-Dependent Cell Cytotoxicity</subject><subject>Antigens, CD20 - immunology</subject><subject>Atomic properties</subject><subject>Binding</subject><subject>CD20 antigen</subject><subject>Cell Line</subject><subject>CHO Cells</subject><subject>Contact potentials</subject><subject>Cricetulus</subject><subject>Crystal structure</subject><subject>Crystallography, X-Ray</subject><subject>Cytotoxicity</subject><subject>Epidermal growth factor</subject><subject>Fucose</subject><subject>Fucose - chemistry</subject><subject>Fucose - genetics</subject><subject>Fucose - immunology</subject><subject>Humans</subject><subject>Immunoglobulin Fc Fragments - chemistry</subject><subject>Immunoglobulin Fc Fragments - genetics</subject><subject>Immunoglobulin Fc Fragments - immunology</subject><subject>Immunoglobulin G</subject><subject>Immunoglobulins</subject><subject>Immunology</subject><subject>Leukemia</subject><subject>Lymphoma</subject><subject>Lymphomas</subject><subject>Models, Molecular</subject><subject>Monoclonal antibodies</subject><subject>Mutants</subject><subject>Mutation</subject><subject>N-glycans</subject><subject>Pharmaceutical sciences</subject><subject>Point Mutation</subject><subject>Polysaccharides</subject><subject>R&D</subject><subject>Receptor mechanisms</subject><subject>Receptors</subject><subject>Receptors, IgG - immunology</subject><subject>Research & development</subject><subject>Research parks</subject><subject>Rituximab</subject><subject>Rituximab - chemistry</subject><subject>Rituximab - genetics</subject><subject>Rituximab - immunology</subject><subject>Targeted cancer therapy</subject><subject>Toxicity</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNptkt9uFCEUxonR2Lr6BkZJvPFmV2CAGW5Mmo3VSZrUVL0mDH-6bGZhBbamz-V7-Eyyu9OmNXIDHH7n4xz4AHiN0QI3Lf6wjrsU1LjYxmAXCFOECXoCTrFoyJwT1Dx9sD4BL3JeI8SajvPn4IRwWkdDT0HuN9uYigrawuhgWVn4zRsLlyvlA1QFfo3ZFx8DJILvibNQ_BDNLTzXsBJ9KDYpvScy_OXLqsb__L7q-15BFQy8rIrpEINXVtttiSm_BM-cGrN9Nc0z8OP80_fll_nF5ed-eXYx14zwMtdW41YxjAbTWd5xg9WgqWJOYK6paY3pDFWOdbylomMdalGj6o4gxURXe52Bt0fd7RiznN4rS9wShDkRlFSiPxImqrXcJr9R6VZG5eUhENO1VKl4PVqJRSsct9QZrWnbuUEPjmmChRNUG8Sr1sfptt2wsUbbUJIaH4k-Pgl-Ja_jjaRMcFY_aQbeTwIp_tzZXOTGZ23HUQUbd4e6ccMY5qKi7_5B_98dPVI6xZyTdffFYCT3HrrLknsPyclDNe3Nw0buk-5M0_wFBMbFWA</recordid><startdate>20151007</startdate><enddate>20151007</enddate><creator>Isoda, Yuya</creator><creator>Yagi, Hirokazu</creator><creator>Satoh, Tadashi</creator><creator>Shibata-Koyama, Mami</creator><creator>Masuda, Kazuhiro</creator><creator>Satoh, Mitsuo</creator><creator>Kato, Koichi</creator><creator>Iida, Shigeru</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20151007</creationdate><title>Importance of the Side Chain at Position 296 of Antibody Fc in Interactions with FcγRIIIa and Other Fcγ Receptors</title><author>Isoda, Yuya ; Yagi, Hirokazu ; Satoh, Tadashi ; Shibata-Koyama, Mami ; Masuda, Kazuhiro ; Satoh, Mitsuo ; Kato, Koichi ; Iida, Shigeru</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c526t-cec17a510bd8e686d1abc4a5f916c4d7dd8d4af5867498580703a58620a598053</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Affinity</topic><topic>Animals</topic><topic>Antibodies</topic><topic>Antibody-Dependent Cell Cytotoxicity</topic><topic>Antigens, CD20 - immunology</topic><topic>Atomic properties</topic><topic>Binding</topic><topic>CD20 antigen</topic><topic>Cell Line</topic><topic>CHO Cells</topic><topic>Contact potentials</topic><topic>Cricetulus</topic><topic>Crystal structure</topic><topic>Crystallography, X-Ray</topic><topic>Cytotoxicity</topic><topic>Epidermal growth factor</topic><topic>Fucose</topic><topic>Fucose - chemistry</topic><topic>Fucose - genetics</topic><topic>Fucose - immunology</topic><topic>Humans</topic><topic>Immunoglobulin Fc Fragments - chemistry</topic><topic>Immunoglobulin Fc Fragments - genetics</topic><topic>Immunoglobulin Fc Fragments - immunology</topic><topic>Immunoglobulin G</topic><topic>Immunoglobulins</topic><topic>Immunology</topic><topic>Leukemia</topic><topic>Lymphoma</topic><topic>Lymphomas</topic><topic>Models, Molecular</topic><topic>Monoclonal antibodies</topic><topic>Mutants</topic><topic>Mutation</topic><topic>N-glycans</topic><topic>Pharmaceutical sciences</topic><topic>Point Mutation</topic><topic>Polysaccharides</topic><topic>R&D</topic><topic>Receptor mechanisms</topic><topic>Receptors</topic><topic>Receptors, IgG - immunology</topic><topic>Research & development</topic><topic>Research parks</topic><topic>Rituximab</topic><topic>Rituximab - chemistry</topic><topic>Rituximab - genetics</topic><topic>Rituximab - immunology</topic><topic>Targeted cancer therapy</topic><topic>Toxicity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Isoda, Yuya</creatorcontrib><creatorcontrib>Yagi, Hirokazu</creatorcontrib><creatorcontrib>Satoh, Tadashi</creatorcontrib><creatorcontrib>Shibata-Koyama, Mami</creatorcontrib><creatorcontrib>Masuda, Kazuhiro</creatorcontrib><creatorcontrib>Satoh, Mitsuo</creatorcontrib><creatorcontrib>Kato, Koichi</creatorcontrib><creatorcontrib>Iida, Shigeru</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Isoda, Yuya</au><au>Yagi, Hirokazu</au><au>Satoh, Tadashi</au><au>Shibata-Koyama, Mami</au><au>Masuda, Kazuhiro</au><au>Satoh, Mitsuo</au><au>Kato, Koichi</au><au>Iida, Shigeru</au><au>Barchi, Joseph J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Importance of the Side Chain at Position 296 of Antibody Fc in Interactions with FcγRIIIa and Other Fcγ Receptors</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2015-10-07</date><risdate>2015</risdate><volume>10</volume><issue>10</issue><spage>e0140120</spage><epage>e0140120</epage><pages>e0140120-e0140120</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Antibody-dependent cellular cytotoxicity (ADCC) is an important effector function determining the clinical efficacy of therapeutic antibodies. Core fucose removal from N-glycans on the Fc portion of immunoglobulin G (IgG) improves the binding affinity for Fcγ receptor IIIa (FcγRIIIa) and dramatically enhances ADCC. Our previous structural analyses revealed that Tyr-296 of IgG1-Fc plays a critical role in the interaction with FcγRIIIa, particularly in the enhanced FcγRIIIa binding of nonfucosylated IgG1. However, the importance of the Tyr-296 residue in the antibody in the interaction with various Fcγ receptors has not yet been elucidated. To further clarify the biological importance of this residue, we established comprehensive Tyr-296 mutants as fucosylated and nonfucosylated anti-CD20 IgG1s rituximab variants and examined their binding to recombinant soluble human Fcγ receptors: shFcγRI, shFcγRIIa, shFcγRIIIa, and shFcγRIIIb. Some of the mutations affected the binding of antibody to not only shFcγRIIIa but also shFcγRIIa and shFcγRIIIb, suggesting that the Tyr-296 residue in the antibody was also involved in interactions with FcγRIIa and FcγRIIIb. For FcγRIIIa binding, almost all Tyr-296 variants showed lower binding affinities than the wild-type antibody, irrespective of their core fucosylation, particularly in Y296K and Y296P. Notably, only the Y296W mutant showed improved binding to FcγRIIIa. The 3.00 Å-resolution crystal structure of the nonfucosylated Y296W mutant in complex with shFcγRIIIa harboring two N-glycans revealed that the Tyr-to-Trp substitution increased the number of potential contact atoms in the complex, thus improving the binding of the antibody to shFcγRIIIa. The nonfucosylated Y296W mutant retained high ADCC activity, relative to the nonfucosylated wild-type IgG1, and showed greater binding affinity for FcγRIIa. Our data may improve our understanding of the biological importance of human IgG1-Fc Tyr-296 in interactions with various Fcγ receptors, and have applications in the modulation of the IgG1-Fc function of therapeutic antibodies.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>26444434</pmid><doi>10.1371/journal.pone.0140120</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2015-10, Vol.10 (10), p.e0140120-e0140120 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1720162942 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Affinity Animals Antibodies Antibody-Dependent Cell Cytotoxicity Antigens, CD20 - immunology Atomic properties Binding CD20 antigen Cell Line CHO Cells Contact potentials Cricetulus Crystal structure Crystallography, X-Ray Cytotoxicity Epidermal growth factor Fucose Fucose - chemistry Fucose - genetics Fucose - immunology Humans Immunoglobulin Fc Fragments - chemistry Immunoglobulin Fc Fragments - genetics Immunoglobulin Fc Fragments - immunology Immunoglobulin G Immunoglobulins Immunology Leukemia Lymphoma Lymphomas Models, Molecular Monoclonal antibodies Mutants Mutation N-glycans Pharmaceutical sciences Point Mutation Polysaccharides R&D Receptor mechanisms Receptors Receptors, IgG - immunology Research & development Research parks Rituximab Rituximab - chemistry Rituximab - genetics Rituximab - immunology Targeted cancer therapy Toxicity |
| title | Importance of the Side Chain at Position 296 of Antibody Fc in Interactions with FcγRIIIa and Other Fcγ Receptors |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-02T20%3A45%3A36IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Importance%20of%20the%20Side%20Chain%20at%20Position%20296%20of%20Antibody%20Fc%20in%20Interactions%20with%20Fc%CE%B3RIIIa%20and%20Other%20Fc%CE%B3%20Receptors&rft.jtitle=PloS%20one&rft.au=Isoda,%20Yuya&rft.date=2015-10-07&rft.volume=10&rft.issue=10&rft.spage=e0140120&rft.epage=e0140120&rft.pages=e0140120-e0140120&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0140120&rft_dat=%3Cproquest_plos_%3E3830531211%3C/proquest_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1720162942&rft_id=info:pmid/26444434&rft_doaj_id=oai_doaj_org_article_1979f6e4fdcc478fbcbf5c219f94cd06&rfr_iscdi=true |