Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation
Identifieur interne : 003169 ( Pmc/Corpus ); précédent : 003168; suivant : 003170Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation
Auteurs : Veli-Matti Lepp Nen ; Denis Tvorogov ; Kaisa Kisko ; Andrea E. Prota ; Michael Jeltsch ; Andrey Anisimov ; Sandra Markovic-Mueller ; Edward Stuttfeld ; Kenneth N. Goldie ; Kurt Ballmer-Hofer ; Kari AlitaloSource :
- Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 2013.
Abstract
Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1–3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4–7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. A thermodynamic analysis of VEGFR-3 deletion mutants showed that D3, D4-5, and D6-7 all contribute to ligand binding. A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs.
Url:
DOI: 10.1073/pnas.1301415110
PubMed: 23878260
PubMed Central: 3740881
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Goldie</name><affiliation><nlm:aff id="aff4">Center for Cellular Imaging and NanoAnalytics, Biozentrum,<institution>University of Basel</institution>, CH-4056 Basel,<country>Switzerland</country></nlm:aff></affiliation></author><author><name sortKey="Ballmer Hofer, Kurt" sort="Ballmer Hofer, Kurt" uniqKey="Ballmer Hofer K" first="Kurt" last="Ballmer-Hofer">Kurt Ballmer-Hofer</name><affiliation><nlm:aff id="aff2">Laboratory of Biomolecular Research,<institution>Paul Scherrer Institute</institution>, CH-5232 Villigen PSI,<country>Switzerland</country>;</nlm:aff></affiliation></author><author><name sortKey="Alitalo, Kari" sort="Alitalo, Kari" uniqKey="Alitalo K" first="Kari" last="Alitalo">Kari Alitalo</name><affiliation><nlm:aff id="aff1">Wihuri Research Institute and Translational Cancer Biology Program, Institute for Molecular Medicine Finland and Helsinki University Central Hospital, Biomedicum Helsinki,<institution>University of Helsinki</institution>, 00014 Helsinki,<country>Finland</country>;</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">23878260</idno><idno type="pmc">3740881</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3740881</idno><idno type="RBID">PMC:3740881</idno><idno type="doi">10.1073/pnas.1301415110</idno><date when="2013">2013</date><idno type="wicri:Area/Pmc/Corpus">003169</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">003169</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation</title><author><name sortKey="Lepp Nen, Veli Matti" sort="Lepp Nen, Veli Matti" uniqKey="Lepp Nen V" first="Veli-Matti" last="Lepp Nen">Veli-Matti Lepp Nen</name><affiliation><nlm:aff id="aff1">Wihuri Research Institute and Translational Cancer Biology Program, Institute for Molecular Medicine Finland and Helsinki University Central Hospital, Biomedicum Helsinki,<institution>University of Helsinki</institution>, 00014 Helsinki,<country>Finland</country>;</nlm:aff></affiliation></author><author><name sortKey="Tvorogov, Denis" sort="Tvorogov, Denis" uniqKey="Tvorogov D" first="Denis" last="Tvorogov">Denis Tvorogov</name><affiliation><nlm:aff id="aff1">Wihuri Research Institute and Translational Cancer Biology Program, Institute for Molecular Medicine Finland and Helsinki University Central Hospital, Biomedicum Helsinki,<institution>University of Helsinki</institution>, 00014 Helsinki,<country>Finland</country>;</nlm:aff></affiliation></author><author><name sortKey="Kisko, Kaisa" sort="Kisko, Kaisa" uniqKey="Kisko K" first="Kaisa" last="Kisko">Kaisa Kisko</name><affiliation><nlm:aff id="aff2">Laboratory of Biomolecular Research,<institution>Paul Scherrer Institute</institution>, CH-5232 Villigen PSI,<country>Switzerland</country>;</nlm:aff></affiliation></author><author><name sortKey="Prota, Andrea E" sort="Prota, Andrea E" uniqKey="Prota A" first="Andrea E." last="Prota">Andrea E. Prota</name><affiliation><nlm:aff id="aff2">Laboratory of Biomolecular Research,<institution>Paul Scherrer Institute</institution>, CH-5232 Villigen PSI,<country>Switzerland</country>;</nlm:aff></affiliation></author><author><name sortKey="Jeltsch, Michael" sort="Jeltsch, Michael" uniqKey="Jeltsch M" first="Michael" last="Jeltsch">Michael Jeltsch</name><affiliation><nlm:aff id="aff1">Wihuri Research Institute and Translational Cancer Biology Program, Institute for Molecular Medicine Finland and Helsinki University Central Hospital, Biomedicum Helsinki,<institution>University of Helsinki</institution>, 00014 Helsinki,<country>Finland</country>;</nlm:aff></affiliation></author><author><name sortKey="Anisimov, Andrey" sort="Anisimov, Andrey" uniqKey="Anisimov A" first="Andrey" last="Anisimov">Andrey Anisimov</name><affiliation><nlm:aff id="aff1">Wihuri Research Institute and Translational Cancer Biology Program, Institute for Molecular Medicine Finland and Helsinki University Central Hospital, Biomedicum Helsinki,<institution>University of Helsinki</institution>, 00014 Helsinki,<country>Finland</country>;</nlm:aff></affiliation></author><author><name sortKey="Markovic Mueller, Sandra" sort="Markovic Mueller, Sandra" uniqKey="Markovic Mueller S" first="Sandra" last="Markovic-Mueller">Sandra Markovic-Mueller</name><affiliation><nlm:aff id="aff2">Laboratory of Biomolecular Research,<institution>Paul Scherrer Institute</institution>, CH-5232 Villigen PSI,<country>Switzerland</country>;</nlm:aff></affiliation></author><author><name sortKey="Stuttfeld, Edward" sort="Stuttfeld, Edward" uniqKey="Stuttfeld E" first="Edward" last="Stuttfeld">Edward Stuttfeld</name><affiliation><nlm:aff id="aff2">Laboratory of Biomolecular Research,<institution>Paul Scherrer Institute</institution>, CH-5232 Villigen PSI,<country>Switzerland</country>;</nlm:aff></affiliation><affiliation><nlm:aff wicri:cut="; and" id="aff3">Structural Biology and Biophysics, Biozentrum, University of Basel, CH-4056 Basel, Switzerland</nlm:aff></affiliation></author><author><name sortKey="Goldie, Kenneth N" sort="Goldie, Kenneth N" uniqKey="Goldie K" first="Kenneth N." last="Goldie">Kenneth N. Goldie</name><affiliation><nlm:aff id="aff4">Center for Cellular Imaging and NanoAnalytics, Biozentrum,<institution>University of Basel</institution>, CH-4056 Basel,<country>Switzerland</country></nlm:aff></affiliation></author><author><name sortKey="Ballmer Hofer, Kurt" sort="Ballmer Hofer, Kurt" uniqKey="Ballmer Hofer K" first="Kurt" last="Ballmer-Hofer">Kurt Ballmer-Hofer</name><affiliation><nlm:aff id="aff2">Laboratory of Biomolecular Research,<institution>Paul Scherrer Institute</institution>, CH-5232 Villigen PSI,<country>Switzerland</country>;</nlm:aff></affiliation></author><author><name sortKey="Alitalo, Kari" sort="Alitalo, Kari" uniqKey="Alitalo K" first="Kari" last="Alitalo">Kari Alitalo</name><affiliation><nlm:aff id="aff1">Wihuri Research Institute and Translational Cancer Biology Program, Institute for Molecular Medicine Finland and Helsinki University Central Hospital, Biomedicum Helsinki,<institution>University of Helsinki</institution>, 00014 Helsinki,<country>Finland</country>;</nlm:aff></affiliation></author></analytic><series><title level="j">Proceedings of the National Academy of Sciences of the United States of America</title><idno type="ISSN">0027-8424</idno><idno type="eISSN">1091-6490</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1–3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4–7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. A thermodynamic analysis of VEGFR-3 deletion mutants showed that D3, D4-5, and D6-7 all contribute to ligand binding. A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Proc Natl Acad Sci U S A</journal-id><journal-id journal-id-type="iso-abbrev">Proc. Natl. Acad. Sci. U.S.A</journal-id><journal-id journal-id-type="hwp">pnas</journal-id><journal-id journal-id-type="pmc">pnas</journal-id><journal-id journal-id-type="publisher-id">PNAS</journal-id><journal-title-group><journal-title>Proceedings of the National Academy of Sciences of the United States of America</journal-title></journal-title-group><issn pub-type="ppub">0027-8424</issn><issn pub-type="epub">1091-6490</issn><publisher><publisher-name>National Academy of Sciences</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">23878260</article-id><article-id pub-id-type="pmc">3740881</article-id><article-id pub-id-type="publisher-id">201301415</article-id><article-id pub-id-type="doi">10.1073/pnas.1301415110</article-id><article-categories><subj-group subj-group-type="heading"><subject>Biological Sciences</subject><subj-group><subject>Biochemistry</subject></subj-group></subj-group></article-categories><title-group><article-title>Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation</article-title><alt-title alt-title-type="short">Mechanisms of VEGFR dimerization and activation</alt-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Leppänen</surname><given-names>Veli-Matti</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref><xref ref-type="author-notes" rid="fn1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Tvorogov</surname><given-names>Denis</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref><xref ref-type="author-notes" rid="fn1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Kisko</surname><given-names>Kaisa</given-names></name><xref ref-type="aff" rid="aff2"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Prota</surname><given-names>Andrea E.</given-names></name><xref ref-type="aff" rid="aff2"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Jeltsch</surname><given-names>Michael</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Anisimov</surname><given-names>Andrey</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Markovic-Mueller</surname><given-names>Sandra</given-names></name><xref ref-type="aff" rid="aff2"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Stuttfeld</surname><given-names>Edward</given-names></name><xref ref-type="aff" rid="aff2"><sup>b</sup></xref><xref ref-type="aff" rid="aff3"><sup>c</sup></xref></contrib><contrib contrib-type="author"><name><surname>Goldie</surname><given-names>Kenneth N.</given-names></name><xref ref-type="aff" rid="aff4"><sup>d</sup></xref></contrib><contrib contrib-type="author"><name><surname>Ballmer-Hofer</surname><given-names>Kurt</given-names></name><xref ref-type="aff" rid="aff2"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Alitalo</surname><given-names>Kari</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref><xref ref-type="corresp" rid="cor1"><sup>2</sup></xref></contrib><aff id="aff1"><sup>a</sup>Wihuri Research Institute and Translational Cancer Biology Program, Institute for Molecular Medicine Finland and Helsinki University Central Hospital, Biomedicum Helsinki,<institution>University of Helsinki</institution>, 00014 Helsinki,<country>Finland</country>;</aff><aff id="aff2"><sup>b</sup>Laboratory of Biomolecular Research,<institution>Paul Scherrer Institute</institution>, CH-5232 Villigen PSI,<country>Switzerland</country>;</aff><aff id="aff3"><sup>c</sup>Structural Biology and Biophysics, Biozentrum, University of Basel, CH-4056 Basel, Switzerland; and</aff><aff id="aff4"><sup>d</sup>Center for Cellular Imaging and NanoAnalytics, Biozentrum,<institution>University of Basel</institution>, CH-4056 Basel,<country>Switzerland</country></aff></contrib-group><author-notes><corresp id="cor1"><sup>2</sup>To whom correspondence should be addressed. E-mail: <email>Kari.Alitalo@helsinki.fi</email>.</corresp><fn fn-type="edited-by"><p>Edited by Napoleone Ferrara, University of California at San Diego, La Jolla, CA, and approved June 26, 2013 (received for review January 23, 2013)</p></fn><fn fn-type="con"><p>Author contributions: V.-M.L., D.T., K.K., M.J., K.N.G., K.B.-H., and K.A. designed research; V.-M.L., D.T., K.K., A.E.P., A.A., S.M.-M., and E.S. performed research; V.-M.L., D.T., K.K., A.E.P., A.A., S.M.-M., and E.S. analyzed data; and V.-M.L., K.B.-H., and K.A. wrote the paper.</p></fn><fn fn-type="equal" id="fn1"><p><sup>1</sup>V.-M.L. and D.T. contributed equally to this work.</p></fn></author-notes><pub-date pub-type="ppub"><day>6</day><month>8</month><year>2013</year></pub-date><pub-date pub-type="epub"><day>22</day><month>7</month><year>2013</year></pub-date><volume>110</volume><issue>32</issue><fpage>12960</fpage><lpage>12965</lpage><self-uri xlink:title="pdf" xlink:type="simple" xlink:href="pnas.201301415.pdf"></self-uri><abstract><p>Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key drivers of blood and lymph vessel formation in development, but also in several pathological processes. VEGF-C signaling through VEGFR-3 promotes lymphangiogenesis, which is a clinically relevant target for treating lymphatic insufficiency and for blocking tumor angiogenesis and metastasis. The extracellular domain of VEGFRs consists of seven Ig homology domains; domains 1–3 (D1-3) are responsible for ligand binding, and the membrane-proximal domains 4–7 (D4-7) are involved in structural rearrangements essential for receptor dimerization and activation. Here we analyzed the crystal structures of VEGF-C in complex with VEGFR-3 domains D1-2 and of the VEGFR-3 D4-5 homodimer. The structures revealed a conserved ligand-binding interface in D2 and a unique mechanism for VEGFR dimerization and activation, with homotypic interactions in D5. Mutation of the conserved residues mediating the D5 interaction (Thr446 and Lys516) and the D7 interaction (Arg737) compromised VEGF-C induced VEGFR-3 activation. A thermodynamic analysis of VEGFR-3 deletion mutants showed that D3, D4-5, and D6-7 all contribute to ligand binding. A structural model of the VEGF-C/VEGFR-3 D1-7 complex derived from small-angle X-ray scattering data is consistent with the homotypic interactions in D5 and D7. Taken together, our data show that ligand-dependent homotypic interactions in D5 and D7 are essential for VEGFR activation, opening promising possibilities for the design of VEGFR-specific drugs.</p></abstract><kwd-group><kwd>signal transduction</kwd><kwd>receptor tyrosine kinase</kwd></kwd-group><counts><page-count count="6"></page-count></counts></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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