A human FVIII inhibitor modulates FVIII surface electrostatics at a VWF‐binding site distant from its epitope
Background: BO2C11 is a human monoclonal factor (F) VIII inhibitor. When bound to the C2 domain of FVIII, the Fab fragment of BO2C11 (FabBO2C11) buries a surface of C2 that contains residues participating in a binding site for von Willebrand factor (VWF). BO2C11 has thus been proposed to neutralize...
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creator | DIMITROV, J. D. ROUMENINA, L. T. PLANTIER, J.‐L. ANDRE, S. SABOULARD, D. MESLIER, Y. PLANCHAIS, C. JACQUEMIN, M. SAINT‐REMY, J.‐M. ATANASOV, B. P. KAVERI, S. V. LACROIX‐DESMAZES, S. |
description | Background: BO2C11 is a human monoclonal factor (F) VIII inhibitor. When bound to the C2 domain of FVIII, the Fab fragment of BO2C11 (FabBO2C11) buries a surface of C2 that contains residues participating in a binding site for von Willebrand factor (VWF). BO2C11 has thus been proposed to neutralize FVIII by steric hindrance. Objectives: The BO2C11 epitope on C2 overlaps with residues located at the periphery of the putative VWF binding site; hence, most of the residues that constitute the VWF binding site on C2 and a3 remain accessible for VWF interaction following BO2C11/FVIII complex formation. We thus investigated the contribution of alternative molecular mechanisms to FVIII inactivation by BO2C11. Methods: Continuum electrostatic calculations were applied to the crystal structure of C2, free or FabBO2C11‐complexed. In silico predictions were confirmed by site‐directed mutagenesis and VWF‐binding assays of the mutated FVIII. Results: Binding of FabBO2C11 to C2 induced perturbations in the electrostatic potential of C2 and in the local electrostatic parameters of 18 charged residues in C2, which are distant from the BO2C11 epitope. Nine of the predicted electrostatic hotspots clustered on the VWF‐binding site of C2. Mutation of some of the predicted electrostatic hotspots has been associated with hemophilia A and reduced VWF binding in vitro. Conclusions: Inhibitors may neutralize FVIII by alteration of protein surface electrostatics at a long distance from their epitope. Perturbation of the electrostatic environment of C2, either upon binding by anti‐FVIII antibodies or consecutive to missense mutations in the F8 gene, may lead to hampered VWF binding and reduced FVIII residence time in circulation. |
doi_str_mv | 10.1111/j.1538-7836.2010.03878.x |
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D. ; ROUMENINA, L. T. ; PLANTIER, J.‐L. ; ANDRE, S. ; SABOULARD, D. ; MESLIER, Y. ; PLANCHAIS, C. ; JACQUEMIN, M. ; SAINT‐REMY, J.‐M. ; ATANASOV, B. P. ; KAVERI, S. V. ; LACROIX‐DESMAZES, S.</creator><creatorcontrib>DIMITROV, J. D. ; ROUMENINA, L. T. ; PLANTIER, J.‐L. ; ANDRE, S. ; SABOULARD, D. ; MESLIER, Y. ; PLANCHAIS, C. ; JACQUEMIN, M. ; SAINT‐REMY, J.‐M. ; ATANASOV, B. P. ; KAVERI, S. V. ; LACROIX‐DESMAZES, S.</creatorcontrib><description>Background: BO2C11 is a human monoclonal factor (F) VIII inhibitor. When bound to the C2 domain of FVIII, the Fab fragment of BO2C11 (FabBO2C11) buries a surface of C2 that contains residues participating in a binding site for von Willebrand factor (VWF). BO2C11 has thus been proposed to neutralize FVIII by steric hindrance. Objectives: The BO2C11 epitope on C2 overlaps with residues located at the periphery of the putative VWF binding site; hence, most of the residues that constitute the VWF binding site on C2 and a3 remain accessible for VWF interaction following BO2C11/FVIII complex formation. We thus investigated the contribution of alternative molecular mechanisms to FVIII inactivation by BO2C11. Methods: Continuum electrostatic calculations were applied to the crystal structure of C2, free or FabBO2C11‐complexed. In silico predictions were confirmed by site‐directed mutagenesis and VWF‐binding assays of the mutated FVIII. Results: Binding of FabBO2C11 to C2 induced perturbations in the electrostatic potential of C2 and in the local electrostatic parameters of 18 charged residues in C2, which are distant from the BO2C11 epitope. Nine of the predicted electrostatic hotspots clustered on the VWF‐binding site of C2. Mutation of some of the predicted electrostatic hotspots has been associated with hemophilia A and reduced VWF binding in vitro. Conclusions: Inhibitors may neutralize FVIII by alteration of protein surface electrostatics at a long distance from their epitope. Perturbation of the electrostatic environment of C2, either upon binding by anti‐FVIII antibodies or consecutive to missense mutations in the F8 gene, may lead to hampered VWF binding and reduced FVIII residence time in circulation.</description><identifier>ISSN: 1538-7933</identifier><identifier>EISSN: 1538-7836</identifier><identifier>DOI: 10.1111/j.1538-7836.2010.03878.x</identifier><identifier>PMID: 20374449</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Antibodies, Monoclonal - immunology ; Antibodies, Monoclonal - pharmacology ; Binding Sites - drug effects ; Epitopes - immunology ; factor VIII ; Factor VIII - antagonists & inhibitors ; Factor VIII - chemistry ; Factor VIII - genetics ; Factor VIII - immunology ; FVIII inhibitors ; Hemophilia A ; Humans ; inhibitory mechanism ; Models, Molecular ; Mutagenesis, Site-Directed ; Mutation, Missense ; Static Electricity ; surface electrostatics ; von Willebrand Factor - metabolism</subject><ispartof>Journal of thrombosis and haemostasis, 2010-07, Vol.8 (7), p.1524-1531</ispartof><rights>2010 International Society on Thrombosis and Haemostasis</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20374449$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>DIMITROV, J. D.</creatorcontrib><creatorcontrib>ROUMENINA, L. T.</creatorcontrib><creatorcontrib>PLANTIER, J.‐L.</creatorcontrib><creatorcontrib>ANDRE, S.</creatorcontrib><creatorcontrib>SABOULARD, D.</creatorcontrib><creatorcontrib>MESLIER, Y.</creatorcontrib><creatorcontrib>PLANCHAIS, C.</creatorcontrib><creatorcontrib>JACQUEMIN, M.</creatorcontrib><creatorcontrib>SAINT‐REMY, J.‐M.</creatorcontrib><creatorcontrib>ATANASOV, B. P.</creatorcontrib><creatorcontrib>KAVERI, S. V.</creatorcontrib><creatorcontrib>LACROIX‐DESMAZES, S.</creatorcontrib><title>A human FVIII inhibitor modulates FVIII surface electrostatics at a VWF‐binding site distant from its epitope</title><title>Journal of thrombosis and haemostasis</title><addtitle>J Thromb Haemost</addtitle><description>Background: BO2C11 is a human monoclonal factor (F) VIII inhibitor. When bound to the C2 domain of FVIII, the Fab fragment of BO2C11 (FabBO2C11) buries a surface of C2 that contains residues participating in a binding site for von Willebrand factor (VWF). BO2C11 has thus been proposed to neutralize FVIII by steric hindrance. Objectives: The BO2C11 epitope on C2 overlaps with residues located at the periphery of the putative VWF binding site; hence, most of the residues that constitute the VWF binding site on C2 and a3 remain accessible for VWF interaction following BO2C11/FVIII complex formation. We thus investigated the contribution of alternative molecular mechanisms to FVIII inactivation by BO2C11. Methods: Continuum electrostatic calculations were applied to the crystal structure of C2, free or FabBO2C11‐complexed. In silico predictions were confirmed by site‐directed mutagenesis and VWF‐binding assays of the mutated FVIII. Results: Binding of FabBO2C11 to C2 induced perturbations in the electrostatic potential of C2 and in the local electrostatic parameters of 18 charged residues in C2, which are distant from the BO2C11 epitope. Nine of the predicted electrostatic hotspots clustered on the VWF‐binding site of C2. Mutation of some of the predicted electrostatic hotspots has been associated with hemophilia A and reduced VWF binding in vitro. Conclusions: Inhibitors may neutralize FVIII by alteration of protein surface electrostatics at a long distance from their epitope. Perturbation of the electrostatic environment of C2, either upon binding by anti‐FVIII antibodies or consecutive to missense mutations in the F8 gene, may lead to hampered VWF binding and reduced FVIII residence time in circulation.</description><subject>Antibodies, Monoclonal - immunology</subject><subject>Antibodies, Monoclonal - pharmacology</subject><subject>Binding Sites - drug effects</subject><subject>Epitopes - immunology</subject><subject>factor VIII</subject><subject>Factor VIII - antagonists & inhibitors</subject><subject>Factor VIII - chemistry</subject><subject>Factor VIII - genetics</subject><subject>Factor VIII - immunology</subject><subject>FVIII inhibitors</subject><subject>Hemophilia A</subject><subject>Humans</subject><subject>inhibitory mechanism</subject><subject>Models, Molecular</subject><subject>Mutagenesis, Site-Directed</subject><subject>Mutation, Missense</subject><subject>Static Electricity</subject><subject>surface electrostatics</subject><subject>von Willebrand Factor - metabolism</subject><issn>1538-7933</issn><issn>1538-7836</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kU1OwzAQhS0EoqVwBeQdqwQndhJnwaKqKA2qxKaUpeXEE-oqf8SOaHccgTNyEhJaOpsZzfv0Fu8hhD3iev3cb10voNyJOA1dn_RfQnnE3d0ZGp-E8_87pnSErozZEuLFgU8u0cgnNGKMxWNUT_GmK2WF5-skSbCuNjrVtm5xWauukBbMUTFdm8sMMBSQ2bY2VlqdGSwtlnj9Nv_5-k51pXT1jo22gJXuicrivK1LrK3B0PS2DVyji1wWBm6Oe4Je54-r2cJZvjwls-nSafw45A7jASgVBcC4UpnKslDmBGIJKfc9xSUlAacyD1QcR0Ga0pT4ISN5FoaEBUxyOkF3B9-mrT86MFaU2mRQFLKCujMiYqFPQ59FPXl7JLu0BCWaVpey3Yv_jHrg4QB86gL2J90jYuhCbMUQsxgiF0MX4q8LsRPPq8Vw0V_C3H5m</recordid><startdate>201007</startdate><enddate>201007</enddate><creator>DIMITROV, J. D.</creator><creator>ROUMENINA, L. T.</creator><creator>PLANTIER, J.‐L.</creator><creator>ANDRE, S.</creator><creator>SABOULARD, D.</creator><creator>MESLIER, Y.</creator><creator>PLANCHAIS, C.</creator><creator>JACQUEMIN, M.</creator><creator>SAINT‐REMY, J.‐M.</creator><creator>ATANASOV, B. P.</creator><creator>KAVERI, S. V.</creator><creator>LACROIX‐DESMAZES, S.</creator><general>Blackwell Publishing Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7T5</scope><scope>H94</scope></search><sort><creationdate>201007</creationdate><title>A human FVIII inhibitor modulates FVIII surface electrostatics at a VWF‐binding site distant from its epitope</title><author>DIMITROV, J. D. ; ROUMENINA, L. T. ; PLANTIER, J.‐L. ; ANDRE, S. ; SABOULARD, D. ; MESLIER, Y. ; PLANCHAIS, C. ; JACQUEMIN, M. ; SAINT‐REMY, J.‐M. ; ATANASOV, B. P. ; KAVERI, S. V. ; LACROIX‐DESMAZES, S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p2968-485edd75e48ddcdcc6af0e9aeb821d8a30583af5d9975bb3b02640fc660454a83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Antibodies, Monoclonal - immunology</topic><topic>Antibodies, Monoclonal - pharmacology</topic><topic>Binding Sites - drug effects</topic><topic>Epitopes - immunology</topic><topic>factor VIII</topic><topic>Factor VIII - antagonists & inhibitors</topic><topic>Factor VIII - chemistry</topic><topic>Factor VIII - genetics</topic><topic>Factor VIII - immunology</topic><topic>FVIII inhibitors</topic><topic>Hemophilia A</topic><topic>Humans</topic><topic>inhibitory mechanism</topic><topic>Models, Molecular</topic><topic>Mutagenesis, Site-Directed</topic><topic>Mutation, Missense</topic><topic>Static Electricity</topic><topic>surface electrostatics</topic><topic>von Willebrand Factor - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>DIMITROV, J. D.</creatorcontrib><creatorcontrib>ROUMENINA, L. T.</creatorcontrib><creatorcontrib>PLANTIER, J.‐L.</creatorcontrib><creatorcontrib>ANDRE, S.</creatorcontrib><creatorcontrib>SABOULARD, D.</creatorcontrib><creatorcontrib>MESLIER, Y.</creatorcontrib><creatorcontrib>PLANCHAIS, C.</creatorcontrib><creatorcontrib>JACQUEMIN, M.</creatorcontrib><creatorcontrib>SAINT‐REMY, J.‐M.</creatorcontrib><creatorcontrib>ATANASOV, B. P.</creatorcontrib><creatorcontrib>KAVERI, S. V.</creatorcontrib><creatorcontrib>LACROIX‐DESMAZES, S.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><jtitle>Journal of thrombosis and haemostasis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>DIMITROV, J. D.</au><au>ROUMENINA, L. T.</au><au>PLANTIER, J.‐L.</au><au>ANDRE, S.</au><au>SABOULARD, D.</au><au>MESLIER, Y.</au><au>PLANCHAIS, C.</au><au>JACQUEMIN, M.</au><au>SAINT‐REMY, J.‐M.</au><au>ATANASOV, B. P.</au><au>KAVERI, S. V.</au><au>LACROIX‐DESMAZES, S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A human FVIII inhibitor modulates FVIII surface electrostatics at a VWF‐binding site distant from its epitope</atitle><jtitle>Journal of thrombosis and haemostasis</jtitle><addtitle>J Thromb Haemost</addtitle><date>2010-07</date><risdate>2010</risdate><volume>8</volume><issue>7</issue><spage>1524</spage><epage>1531</epage><pages>1524-1531</pages><issn>1538-7933</issn><eissn>1538-7836</eissn><abstract>Background: BO2C11 is a human monoclonal factor (F) VIII inhibitor. When bound to the C2 domain of FVIII, the Fab fragment of BO2C11 (FabBO2C11) buries a surface of C2 that contains residues participating in a binding site for von Willebrand factor (VWF). BO2C11 has thus been proposed to neutralize FVIII by steric hindrance. Objectives: The BO2C11 epitope on C2 overlaps with residues located at the periphery of the putative VWF binding site; hence, most of the residues that constitute the VWF binding site on C2 and a3 remain accessible for VWF interaction following BO2C11/FVIII complex formation. We thus investigated the contribution of alternative molecular mechanisms to FVIII inactivation by BO2C11. Methods: Continuum electrostatic calculations were applied to the crystal structure of C2, free or FabBO2C11‐complexed. In silico predictions were confirmed by site‐directed mutagenesis and VWF‐binding assays of the mutated FVIII. Results: Binding of FabBO2C11 to C2 induced perturbations in the electrostatic potential of C2 and in the local electrostatic parameters of 18 charged residues in C2, which are distant from the BO2C11 epitope. Nine of the predicted electrostatic hotspots clustered on the VWF‐binding site of C2. Mutation of some of the predicted electrostatic hotspots has been associated with hemophilia A and reduced VWF binding in vitro. Conclusions: Inhibitors may neutralize FVIII by alteration of protein surface electrostatics at a long distance from their epitope. Perturbation of the electrostatic environment of C2, either upon binding by anti‐FVIII antibodies or consecutive to missense mutations in the F8 gene, may lead to hampered VWF binding and reduced FVIII residence time in circulation.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>20374449</pmid><doi>10.1111/j.1538-7836.2010.03878.x</doi><tpages>8</tpages></addata></record> |
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subjects | Antibodies, Monoclonal - immunology Antibodies, Monoclonal - pharmacology Binding Sites - drug effects Epitopes - immunology factor VIII Factor VIII - antagonists & inhibitors Factor VIII - chemistry Factor VIII - genetics Factor VIII - immunology FVIII inhibitors Hemophilia A Humans inhibitory mechanism Models, Molecular Mutagenesis, Site-Directed Mutation, Missense Static Electricity surface electrostatics von Willebrand Factor - metabolism |
title | A human FVIII inhibitor modulates FVIII surface electrostatics at a VWF‐binding site distant from its epitope |
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