PMID- 28827556 OWN - NLM STAT- MEDLINE DCOM- 20190403 LR - 20211204 IS - 2045-2322 (Electronic) IS - 2045-2322 (Linking) VI - 7 IP - 1 DP - 2017 Aug 21 TI - Characterization and structural determination of a new anti-MET function-blocking antibody with binding epitope distinct from the ligand binding domain. PG - 9000 LID - 10.1038/s41598-017-09460-2 [doi] LID - 9000 AB - The growth and motility factor Hepatocyte Growth Factor/Scatter Factor (HGF/SF) and its receptor, the product of the MET proto-oncogene, promote invasion and metastasis of tumor cells and have been considered potential targets for cancer therapy. We generated a new Met-blocking antibody which binds outside the ligand-binding site, and determined the crystal structure of the Fab in complex with its target, which identifies the binding site as the Met Ig1 domain. The antibody, 107_A07, inhibited HGF/SF-induced cell migration and proliferation in vitro and inhibited growth of tumor xenografts in vivo. In biochemical assays, 107_A07 competes with both HGF/SF and its truncated splice variant NK1 for MET binding, despite the location of the antibody epitope on a domain (Ig1) not reported to bind NK1 or HGF/SF. Overlay of the Fab-MET crystal structure with the InternalinB-MET crystal structure shows that the 107_A07 Fab comes into close proximity with the HGF/SF-binding SEMA domain when MET is in the "compact", InternalinB-bound conformation, but not when MET is in the "open" conformation. These findings provide further support for the importance of the "compact" conformation of the MET extracellular domain, and the relevance of this conformation to HGF/SF binding and signaling. FAU - DiCara, Danielle M AU - DiCara DM AD - MRC Centre, Hills Road, Cambridge, CB2 2QH, UK. AD - Department of Oncology, University of Cambridge, Cambridge Biomedical Campus, Cambridge, CB2 0XZ, UK. AD - Genentech Inc., South San Francisco, 94080, USA. FAU - Chirgadze, Dimitri Y AU - Chirgadze DY AD - Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK. FAU - Pope, Anthony R AU - Pope AR AD - IONTAS Ltd, Babraham Institute, Babraham, Cambridgeshire, CB22 3AT, UK. FAU - Karatt-Vellatt, Aneesh AU - Karatt-Vellatt A AD - IONTAS Ltd, Babraham Institute, Babraham, Cambridgeshire, CB22 3AT, UK. FAU - Winter, Anja AU - Winter A AUID- ORCID: 0000-0002-8602-8009 AD - Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK. AD - Faculty of Natural Sciences, Keele University, Staffordshire, ST5 5BG, UK. FAU - Slavny, Peter AU - Slavny P AD - IONTAS Ltd, Babraham Institute, Babraham, Cambridgeshire, CB22 3AT, UK. FAU - van den Heuvel, Joop AU - van den Heuvel J AUID- ORCID: 0000-0001-5085-4010 AD - Helmholtz Zentrum fur Infektionsforschung, Inhoffenstrasse 7, 38124, Braunschweig, Germany. FAU - Parthiban, Kothai AU - Parthiban K AD - IONTAS Ltd, Babraham Institute, Babraham, Cambridgeshire, CB22 3AT, UK. FAU - Holland, Jane AU - Holland J AD - Max Delbrueck Center for Molecular Medicine (MDC) in the Helmholtz Association, 13125, Berlin, Germany. FAU - Packman, Len C AU - Packman LC AD - Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK. FAU - Mavria, Georgia AU - Mavria G AD - Leeds Institute of Cancer and Pathology, University of Leeds, St James' University Hospital, Beckett Street, Leeds, LS9 7TF, UK. FAU - Hoffmann, Jens AU - Hoffmann J AD - Experimental Pharmacology & Oncology Berlin-Buch GmbH, Robert-Rossle-Str. 10, 13125, Berlin-Buch, Germany. FAU - Birchmeier, Walter AU - Birchmeier W AD - Max Delbrueck Center for Molecular Medicine (MDC) in the Helmholtz Association, 13125, Berlin, Germany. FAU - Gherardi, Ermanno AU - Gherardi E AD - MRC Centre, Hills Road, Cambridge, CB2 2QH, UK. egherard@unipv.it. AD - Department of Oncology, University of Cambridge, Cambridge Biomedical Campus, Cambridge, CB2 0XZ, UK. egherard@unipv.it. AD - Division of Immunology and General Pathology, Department of Molecular Medicine, 1 via A Ferrata, 27100, Pavia, Italy. egherard@unipv.it. FAU - McCafferty, John AU - McCafferty J AD - Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK. jmc@iontas.co.uk. AD - IONTAS Ltd, Babraham Institute, Babraham, Cambridgeshire, CB22 3AT, UK. jmc@iontas.co.uk. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20170821 PL - England TA - Sci Rep JT - Scientific reports JID - 101563288 RN - 0 (Antibodies, Blocking) RN - 0 (Antineoplastic Agents, Immunological) RN - 0 (Immunoglobulin Fab Fragments) RN - 0 (MAS1 protein, human) RN - 0 (Proto-Oncogene Mas) RN - EC 2.7.10.1 (MET protein, human) RN - EC 2.7.10.1 (Proto-Oncogene Proteins c-met) SB - IM MH - Animals MH - Antibodies, Blocking/administration & dosage/chemistry/*isolation & purification/*metabolism MH - Antineoplastic Agents, Immunological/administration & dosage/chemistry/*isolation & purification/*metabolism MH - Binding Sites MH - Cell Line, Tumor MH - Cell Movement/drug effects MH - Cell Proliferation/drug effects MH - Crystallography, X-Ray MH - Disease Models, Animal MH - Glioblastoma/drug therapy MH - Heterografts MH - Humans MH - Immunoglobulin Fab Fragments/administration & dosage/chemistry/isolation & purification/metabolism MH - Mice, Nude MH - Neoplasm Transplantation MH - Protein Binding MH - Protein Conformation MH - Proto-Oncogene Mas MH - Proto-Oncogene Proteins c-met/*metabolism MH - Treatment Outcome PMC - PMC5567289 COIS- The authors declare that they have no competing interests. EDAT- 2017/08/23 06:00 MHDA- 2019/04/04 06:00 PMCR- 2017/08/21 CRDT- 2017/08/23 06:00 PHST- 2017/05/08 00:00 [received] PHST- 2017/07/25 00:00 [accepted] PHST- 2017/08/23 06:00 [entrez] PHST- 2017/08/23 06:00 [pubmed] PHST- 2019/04/04 06:00 [medline] PHST- 2017/08/21 00:00 [pmc-release] AID - 10.1038/s41598-017-09460-2 [pii] AID - 9460 [pii] AID - 10.1038/s41598-017-09460-2 [doi] PST - epublish SO - Sci Rep. 2017 Aug 21;7(1):9000. doi: 10.1038/s41598-017-09460-2.