IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Structure of the human receptor tyrosine kinase met in complex with the Listeria invasion protein InlB. Structure of the human receptor tyrosine kinase met in complex with the Listeria invasion protein InlB. Cell (0092-8674) 2007 imex curation Niemann HH., Jager V., Butler PJ., van den Heuvel J., Schmidt S., Ferraris D., Gherardi E., Heinz DW. Monomeric Met constructs were bound to Ni-coated HisGrab plates (Pierce) via their C-terminal His-tag. HRP-coupled anti-GST antibody was used for detection with an ABTS staining reaction. egherard@mrc-lmb.cam.ac.uk 07-Sept-2007: Contacted by JYOTI. davet Only protein-protein interactions 4 in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Niemann HH., Jager V., Butler PJ., van den Heuvel J., Schmidt S., Ferraris D., Gherardi E., Heinz DW. Monomeric Met constructs were bound to Ni-coated HisGrab plates (Pierce) via their C-terminal His-tag. HRP-coupled anti-GST antibody was used for detection with an ABTS staining reaction. egherard@mrc-lmb.cam.ac.uk egherard@mrc-lmb.cam.ac.uk Accepted 2007-SEP-8 by DAVET. 07-Sept-2007: Contacted by JYOTI. Cell (0092-8674) 2007 davet Only protein-protein interactions 4 imex curation Structure of the human receptor tyrosine kinase met in complex with the Listeria invasion protein InlB. Structure of the human receptor tyrosine kinase met in complex with the Listeria invasion protein InlB. Cell (0092-8674) 2007 imex curation Niemann HH., Jager V., Butler PJ., van den Heuvel J., Schmidt S., Ferraris D., Gherardi E., Heinz DW. Monomeric Met constructs were bound to Ni-coated HisGrab plates (Pierce) via their C-terminal His-tag. HRP-coupled anti-GST antibody was used for detection with an ABTS staining reaction. egherard@mrc-lmb.cam.ac.uk 07-Sept-2007: Contacted by JYOTI. davet Only protein-protein interactions 4 in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa predetermined predetermined participant predetermined Niemann HH., Jager V., Butler PJ., van den Heuvel J., Schmidt S., Ferraris D., Gherardi E., Heinz DW. Monomeric Met constructs were bound to Ni-coated HisGrab plates (Pierce) via their C-terminal His-tag. HRP-coupled anti-GST antibody was used for detection with an ABTS staining reaction. egherard@mrc-lmb.cam.ac.uk egherard@mrc-lmb.cam.ac.uk Accepted 2007-SEP-8 by DAVET. 07-Sept-2007: Contacted by JYOTI. Cell (0092-8674) 2007 davet Only protein-protein interactions 4 imex curation Structure of the human receptor tyrosine kinase met in complex with the Listeria invasion protein InlB. Structure of the human receptor tyrosine kinase met in complex with the Listeria invasion protein InlB. Cell (0092-8674) 2007 imex curation Niemann HH., Jager V., Butler PJ., van den Heuvel J., Schmidt S., Ferraris D., Gherardi E., Heinz DW. Monomeric Met constructs were bound to Ni-coated HisGrab plates (Pierce) via their C-terminal His-tag. HRP-coupled anti-GST antibody was used for detection with an ABTS staining reaction. egherard@mrc-lmb.cam.ac.uk 07-Sept-2007: Contacted by JYOTI. davet Only protein-protein interactions 4 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Niemann HH., Jager V., Butler PJ., van den Heuvel J., Schmidt S., Ferraris D., Gherardi E., Heinz DW. Monomeric Met constructs were bound to Ni-coated HisGrab plates (Pierce) via their C-terminal His-tag. HRP-coupled anti-GST antibody was used for detection with an ABTS staining reaction. egherard@mrc-lmb.cam.ac.uk egherard@mrc-lmb.cam.ac.uk Accepted 2007-SEP-8 by DAVET. 07-Sept-2007: Contacted by JYOTI. Cell (0092-8674) 2007 davet Only protein-protein interactions 4 imex curation met_human Hepatocyte growth factor receptor MET Scatter factor receptor HGF/SF receptor Tyrosine-protein kinase Met Proto-oncogene c-Met protein protein human Homo sapiens Human MKAPAVLAPGILVLLFTLVQRSNGECKEALAKSEMNVNMKYQLPNFTAETPIQNVILHEHHIFLGATNYIYVLNEEDLQKVAEYKTGPVLEHPDCFPCQDCSSKANLSGGVWKDNINMALVVDTYYDDQLISCGSVNRGTCQRHVFPHNHTADIQSEVHCIFSPQIEEPSQCPDCVVSALGAKVLSSVKDRFINFFVGNTINSSYFPDHPLHSISVRRLKETKDGFMFLTDQSYIDVLPEFRDSYPIKYVHAFESNNFIYFLTVQRETLDAQTFHTRIIRFCSINSGLHSYMEMPLECILTEKRKKRSTKKEVFNILQAAYVSKPGAQLARQIGASLNDDILFGVFAQSKPDSAEPMDRSAMCAFPIKYVNDFFNKIVNKNNVRCLQHFYGPNHEHCFNRTLLRNSSGCEARRDEYRTEFTTALQRVDLFMGQFSEVLLTSISTFIKGDLTIANLGTSEGRFMQVVVSRSGPSTPHVNFLLDSHPVSPEVIVEHTLNQNGYTLVITGKKITKIPLNGLGCRHFQSCSQCLSAPPFVQCGWCHDKCVRSEECLSGTWTQQICLPAIYKVFPNSAPLEGGTRLTICGWDFGFRRNNKFDLKKTRVLLGNESCTLTLSESTMNTLKCTVGPAMNKHFNMSIIISNGHGTTQYSTFSYVDPVITSISPKYGPMAGGTLLTLTGNYLNSGNSRHISIGGKTCTLKSVSNSILECYTPAQTISTEFAVKLKIDLANRETSIFSYREDPIVYEIHPTKSFISGGSTITGVGKNLNSVSVPRMVINVHEAGRNFTVACQHRSNSEIICCTTPSLQQLNLQLPLKTKAFFMLDGILSKYFDLIYVHNPVFKPFEKPVMISMGNENVLEIKGNDIDPEAVKGEVLKVGNKSCENIHLHSEAVLCTVPNDLLKLNSELNIEWKQAISSTVLGKVIVQPDQNFTGLIAGVVSISTALLLLLGFFLWLKKRKQIKDLGSELVRYDARVHTPHLDRLVSARSVSPTTEMVSNESVDYRATFPEDQFPNSSQNGSCRQVQYPLTDMSPILTSGDSDISSPLLQNTVHIDLSALNPELVQAVQHVVIGPSSLIVHFNEVIGRGHFGCVYHGTLLDNDGKKIHCAVKSLNRITDIGEVSQFLTEGIIMKDFSHPNVLSLLGICLRSEGSPLVVLPYMKHGDLRNFIRNETHNPTVKDLIGFGLQVAKGMKYLASKKFVHRDLAARNCMLDEKFTVKVADFGLARDMYDKEYYSVHNKTGAKLPVKWMALESLQTQKFTTKSDVWSFGVLLWELMTRGAPPYPDVNTFDITVYLLQGRRLLQPEYCPDPLYEVMLKCWHPKAEMRPSFSELVSRISAIFSTFIGEHYVHVNATYVNVKCVAPYPSLLSSEDNADDEVDTRPASFWETS DIP protein P08581 original sequence version: 94 DIP protein P08581 original sequence version: 200 DIP protein P08581 original sequence version: 128 DIP protein P08581-1 original sequence version: 166 DIP protein P08581 original sequence version: 211 9CF896D1273905C3 inlb_lismo Internalin B inlB Invasion protein InlB lmo0434 protein protein lismo Listeria monocytogenes serovar 1/2a (strain ATCC BAA-679 / EGD-e) MKEKHNPRRKYCLISGLAIIFSLWIIIGNGAKVQAETITVPTPIKQIFSDDAFAETIKDNLKKKSVTDAVTQNELNSIDQIIANNSDIKSVQGIQYLPNVTKLFLNGNKLTDIKPLANLKNLGWLFLDENKVKDLSSLKDLKKLKSLSLEHNGISDINGLVHLPQLESLYLGNNKITDITVLSRLTKLDTLSLEDNQISDIVPLAGLTKLQNLYLSKNHISDLRALAGLKNLDVLELFSQECLNKPINHQSNLVVPNTVKNTDGSLVTPEIISDDGDYEKPNVKWHLPEFTNEVSFIFYQPVTIGKAKARFHGRVTQPLKEVYTVSYDVDGTVIKTKVEAGTRITAPKPPTKQGYVFKGWYTEKNGGHEWNFNTDYMSGNDFTLYAVFKAETTEKAVNLTRYVKYIRGNAGIYKLPREDNSLKQGTLASHRCKALTVDREARNGGKLWYRLKNIGWTKAENLSLDRYDKMEYDKGVTAYARVRNASGNSVWTKPYNTAGAKHVNKLSVYQGKNMRILREAKTPITTWYQFSIGGKVIGWVDTRALNTFYKQSMEKPTRLTRYVSANKAGESYYKVPVADNPVKRGTLAKYKNQKLIVDCQATIEGQLWYRIRTSSTFIGWTKAANLRAQK CA05F31BEFE9748C heparin Heparin small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) met-inlb-1 2 5 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role bait bait c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. 13 9 direct interaction direct interaction In contrast, InlB321 bound with high- and virtually identical affinity to longer variants of the Met ectodomain (Met741, Met838, and Met928) that contain two, three, or four immunoglobulin (Ig)-like domains, respectively (Figure 1F). Figure 1E & 1F met-inlb-2 3 4 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain met-600 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain met-599 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain tyr-170 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain tyr-170 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 18 23 22 19 direct interaction direct interaction We present the crystal structure of the complex between a large fragment of the human Met ectodomain and the Met-binding domain of InlB. Figure 2 & Supp. Table 1 2.8A (Deglycosylated), 4.0 (glycosylated) A contact between the concave face of the InlB LRR region and Met Ig1 forms the primary interface, the secondary, less extensive contact involves the InlB IR region and the Sema domain of Met. met-inlb-3 1 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. direct interaction direct interaction Velocity sedimentation analysis of the Met928-InlB321 complex Figure 6 Equimolar mixtures of InlB321 and Met928 produced a 1:1 complex. met-inlb-4 1 6 unspecified role unspecified role neutral component neutral component 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role neutral component neutral component region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. direct interaction direct interaction Velocity sedimentation analysis of the Met928-InlBfl complex (2 μM) with a 2-fold excess of dp12 heparin. Figure 6 Heparin induced massive clustering of the Met-InlBfl complex, which shifted to large sedimentation coefficients. met-inlb-5 2 5 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role bait bait c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. 41 45 direct interaction direct interaction InlB241 and InlB321 bind to Met928 with identical apparent affinity in a solid phase binding assay. Figure 5A