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/ http://www.ebi.ac.uk/ cecchi-2012-2 Targeted disruption of heparan sulfate interaction with hepatocyte and vascular endothelial growth factors blocks normal and oncogenic signaling. in vitro In vitro crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Cecchi F., Pajalunga D., Fowler C.A., Uren A., Rabe D.C., Peruzzi B., Macdonald N.J., Blackman D.K., Stahl S.J., Byrd R.A., Bottaro D.P. Cancer - Interactions investigated in the context of cancer Cross linking agent:- bis(sulfosuccinimidyl) suberate. NRP1-Fc was detected by an anti-tag antibody. The authors confirm that 'VEGF' corresponds to GenBank accession NP_001165097 (P15692-4);none of the recombinant NK1 WT or mutant proteins used contain the signal sequence; both the Met and KDR fusion proteins are Human in origin (KDR is from R+D Systems Cat no 357-KD, Met was from R+D Systems Cat no 358-MT, as well as from Genentech (Cat no unknown)- both proteins behave identically in the assays described here). [Amino acid numbering for VEGF165 mutants includes the 26 aa signal sequence, while the publication uses numbering from the mature protein.] dbottaro@helix.nih.gov Cancer Cell (1535-6108) 2012 Only protein-protein interactions imex curation cecchi-2012-1 Targeted disruption of heparan sulfate interaction with hepatocyte and vascular endothelial growth factors blocks normal and oncogenic signaling. in vitro In vitro saturation binding saturation binding predetermined predetermined participant predetermined Cecchi F., Pajalunga D., Fowler C.A., Uren A., Rabe D.C., Peruzzi B., Macdonald N.J., Blackman D.K., Stahl S.J., Byrd R.A., Bottaro D.P. Cancer - Interactions investigated in the context of cancer Cross linking agent:- bis(sulfosuccinimidyl) suberate. NRP1-Fc was detected by an anti-tag antibody. The authors confirm that 'VEGF' corresponds to GenBank accession NP_001165097 (P15692-4);none of the recombinant NK1 WT or mutant proteins used contain the signal sequence; both the Met and KDR fusion proteins are Human in origin (KDR is from R+D Systems Cat no 357-KD, Met was from R+D Systems Cat no 358-MT, as well as from Genentech (Cat no unknown)- both proteins behave identically in the assays described here). [Amino acid numbering for VEGF165 mutants includes the 26 aa signal sequence, while the publication uses numbering from the mature protein.] dbottaro@helix.nih.gov Cancer Cell (1535-6108) 2012 Only protein-protein interactions imex curation cecchi-2012-8 Targeted disruption of heparan sulfate interaction with hepatocyte and vascular endothelial growth factors blocks normal and oncogenic signaling. in vitro In vitro crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Cecchi F., Pajalunga D., Fowler C.A., Uren A., Rabe D.C., Peruzzi B., Macdonald N.J., Blackman D.K., Stahl S.J., Byrd R.A., Bottaro D.P. Cancer - Interactions investigated in the context of cancer Cross linking agent:- bis(sulfosuccinimidyl) suberate. NRP1-Fc was detected by an anti-tag antibody. The authors confirm that 'VEGF' corresponds to GenBank accession NP_001165097 (P15692-4);none of the recombinant NK1 WT or mutant proteins used contain the signal sequence; both the Met and KDR fusion proteins are Human in origin (KDR is from R+D Systems Cat no 357-KD, Met was from R+D Systems Cat no 358-MT, as well as from Genentech (Cat no unknown)- both proteins behave identically in the assays described here). [Amino acid numbering for VEGF165 mutants includes the 26 aa signal sequence, while the publication uses numbering from the mature protein.] dbottaro@helix.nih.gov Cancer Cell (1535-6108) 2012 Only protein-protein interactions imex curation cecchi-2012-3 Targeted disruption of heparan sulfate interaction with hepatocyte and vascular endothelial growth factors blocks normal and oncogenic signaling. human-ht_29 Homo sapiens HT-29 caucasian women with colon adenocarcinoma ht_29 HT-29 confirmed as human with IEF of AST, LDH established from the primary tumor of a 44-year-old Caucasian woman with colon adenocarcinoma in 1964; described to be heterotransplantable forming well-differentiated grade I tumors crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Cecchi F., Pajalunga D., Fowler C.A., Uren A., Rabe D.C., Peruzzi B., Macdonald N.J., Blackman D.K., Stahl S.J., Byrd R.A., Bottaro D.P. Cancer - Interactions investigated in the context of cancer Cross linking agent:- bis(sulfosuccinimidyl) suberate. NRP1-Fc was detected by an anti-tag antibody. The authors confirm that 'VEGF' corresponds to GenBank accession NP_001165097 (P15692-4);none of the recombinant NK1 WT or mutant proteins used contain the signal sequence; both the Met and KDR fusion proteins are Human in origin (KDR is from R+D Systems Cat no 357-KD, Met was from R+D Systems Cat no 358-MT, as well as from Genentech (Cat no unknown)- both proteins behave identically in the assays described here). [Amino acid numbering for VEGF165 mutants includes the 26 aa signal sequence, while the publication uses numbering from the mature protein.] dbottaro@helix.nih.gov Cancer Cell (1535-6108) 2012 Only protein-protein interactions imex curation cecchi-2012-4 Targeted disruption of heparan sulfate interaction with hepatocyte and vascular endothelial growth factors blocks normal and oncogenic signaling. human-pc3m Human PC3M Caucasian prostate adenocarcinoma pc3m Human PC3M Caucasian prostate adenocarcinoma PC-3M was isolated from a liver metastasis produced by the intrasplenic injection of human PC-3 prostate cancer cells into nude mice. PC-3M cells have "Hyperactive Akt" and are PTEN null. crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Cecchi F., Pajalunga D., Fowler C.A., Uren A., Rabe D.C., Peruzzi B., Macdonald N.J., Blackman D.K., Stahl S.J., Byrd R.A., Bottaro D.P. Cancer - Interactions investigated in the context of cancer Cross linking agent:- bis(sulfosuccinimidyl) suberate. NRP1-Fc was detected by an anti-tag antibody. The authors confirm that 'VEGF' corresponds to GenBank accession NP_001165097 (P15692-4);none of the recombinant NK1 WT or mutant proteins used contain the signal sequence; both the Met and KDR fusion proteins are Human in origin (KDR is from R+D Systems Cat no 357-KD, Met was from R+D Systems Cat no 358-MT, as well as from Genentech (Cat no unknown)- both proteins behave identically in the assays described here). [Amino acid numbering for VEGF165 mutants includes the 26 aa signal sequence, while the publication uses numbering from the mature protein.] dbottaro@helix.nih.gov Cancer Cell (1535-6108) 2012 Only protein-protein interactions imex curation cecchi-2012-6 Targeted disruption of heparan sulfate interaction with hepatocyte and vascular endothelial growth factors blocks normal and oncogenic signaling. in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel western blot western blot Immuno blot Immuno blot Cecchi F., Pajalunga D., Fowler C.A., Uren A., Rabe D.C., Peruzzi B., Macdonald N.J., Blackman D.K., Stahl S.J., Byrd R.A., Bottaro D.P. Cancer - Interactions investigated in the context of cancer Cross linking agent:- bis(sulfosuccinimidyl) suberate. NRP1-Fc was detected by an anti-tag antibody. The authors confirm that 'VEGF' corresponds to GenBank accession NP_001165097 (P15692-4);none of the recombinant NK1 WT or mutant proteins used contain the signal sequence; both the Met and KDR fusion proteins are Human in origin (KDR is from R+D Systems Cat no 357-KD, Met was from R+D Systems Cat no 358-MT, as well as from Genentech (Cat no unknown)- both proteins behave identically in the assays described here). [Amino acid numbering for VEGF165 mutants includes the 26 aa signal sequence, while the publication uses numbering from the mature protein.] dbottaro@helix.nih.gov Cancer Cell (1535-6108) 2012 Only protein-protein interactions imex curation cecchi-2012-9 Targeted disruption of heparan sulfate interaction with hepatocyte and vascular endothelial growth factors blocks normal and oncogenic signaling. in vitro In vitro affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom predetermined predetermined participant predetermined Cecchi F., Pajalunga D., Fowler C.A., Uren A., Rabe D.C., Peruzzi B., Macdonald N.J., Blackman D.K., Stahl S.J., Byrd R.A., Bottaro D.P. Cancer - Interactions investigated in the context of cancer Cross linking agent:- bis(sulfosuccinimidyl) suberate. NRP1-Fc was detected by an anti-tag antibody. The authors confirm that 'VEGF' corresponds to GenBank accession NP_001165097 (P15692-4);none of the recombinant NK1 WT or mutant proteins used contain the signal sequence; both the Met and KDR fusion proteins are Human in origin (KDR is from R+D Systems Cat no 357-KD, Met was from R+D Systems Cat no 358-MT, as well as from Genentech (Cat no unknown)- both proteins behave identically in the assays described here). [Amino acid numbering for VEGF165 mutants includes the 26 aa signal sequence, while the publication uses numbering from the mature protein.] dbottaro@helix.nih.gov Cancer Cell (1535-6108) 2012 Only protein-protein interactions imex curation p14210-6 Hepatocyte growth factor HGF/NK1 HGF HPTA Hepatopoietin-A Scatter factor protein protein human Homo sapiens Human MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPALKIKTKKVNTADQCANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGHEFDLYENKDYIRNCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGKDLQENYCRNPRGEEGGPWCFTSNPEVRYEVCDIPQCSEGK p15692-4 Vascular endothelial growth factor A VEGF VEGFA VEGF Vascular permeability factor protein protein human Homo sapiens Human MNFLLSWVHWSLALLLYLHHAKWSQAAPMAEGGGQNHHEVVKFMDVYQRSYCHPIETLVDIFQEYPDEIEYIFKPSCVPLMRCGGCCNDEGLECVPTEESNITMQIMRIKPHQGQHIGEMSFLQHNKCECRPKKDRARQENPCGPCSERRKHLFVQDPQTCKCSCKNTDSRCKARQLELNERTCRCDKPRR o14786-2 Neuropilin-1 Soluble SNRP1 NRP1 NRP VEGF165R Vascular endothelial cell growth factor 165 receptor protein protein human Homo sapiens Human MERGLPLLCAVLALVLAPAGAFRNDKCGDTIKIESPGYLTSPGYPHSYHPSEKCEWLIQAPDPYQRIMINFNPHFDLEDRDCKYDYVEVFDGENENGHFRGKFCGKIAPPPVVSSGPFLFIKFVSDYETHGAGFSIRYEIFKRGPECSQNYTTPSGVIKSPGFPEKYPNSLECTYIVFVPKMSEIILEFESFDLEPDSNPPGGMFCRYDRLEIWDGFPDVGPHIGRYCGQKTPGRIRSSSGILSMVFYTDSAIAKEGFSANYSVLQSSVSEDFKCMEALGMESGEIHSDQITASSQYSTNWSAERSRLNYPENGWTPGEDSYREWIQVDLGLLRFVTAVGTQGAISKETKKKYYVKTYKIDVSSNGEDWITIKEGNKPVLFQGNTNPTDVVVAVFPKPLITRFVRIKPATWETGISMRFEVYGCKITDYPCSGMLGMVSGLISDSQITSSNQGDRNWMPENIRLVTSRSGWALPPAPHSYINEWLQIDLGEEKIVRGIIIQGGKHRENKVFMRKFKIGYSNNGSDWKMIMDDSKRKAKSFEGNNNYDTPELRTFPALSTRFIRIYPERATHGGLGLRMELLGCEVEAPTAGPTTPNGNLVDECDDDQANCHSGTGDDFQLTGGTTVLATEKPTVIDSTIQSGIK 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 heparan_sulfate Heparan_Sulfate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) heparin Heparin small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) vgfr2_human Vascular endothelial growth factor receptor 2 Kinase insert domain receptor Protein-tyrosine kinase receptor flk-1 KDR FLK1 Fetal liver kinase 1 VEGFR2 protein protein human Homo sapiens Human MQSKVLLAVALWLCVETRAASVGLPSVSLDLPRLSIQKDILTIKANTTLQITCRGQRDLDWLWPNNQSGSEQRVEVTECSDGLFCKTLTIPKVIGNDTGAYKCFYRETDLASVIYVYVQDYRSPFIASVSDQHGVVYITENKNKTVVIPCLGSISNLNVSLCARYPEKRFVPDGNRISWDSKKGFTIPSYMISYAGMVFCEAKINDESYQSIMYIVVVVGYRIYDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKPFVAFGSGMESLVEATVGERVRIPAKYLGYPPPEIKWYKNGIPLESNHTIKAGHVLTIMEVSERDTGNYTVILTNPISKEKQSHVVSLVVYVPPQIGEKSLISPVDSYQYGTTQTLTCTVYAIPPPHHIHWYWQLEEECANEPSQAVSVTNPYPCEEWRSVEDFQGGNKIEVNKNQFALIEGKNKTVSTLVIQAANVSALYKCEAVNKVGRGERVISFHVTRGPEITLQPDMQPTEQESVSLWCTADRSTFENLTWYKLGPQPLPIHVGELPTPVCKNLDTLWKLNATMFSNSTNDILIMELKNASLQDQGDYVCLAQDRKTKKRHCVVRQLTVLERVAPTITGNLENQTTSIGESIEVSCTASGNPPPQIMWFKDNETLVEDSGIVLKDGNRNLTIRRVRKEDEGLYTCQACSVLGCAKVEAFFIIEGAQEKTNLEIIILVGTAVIAMFFWLLLVIILRTVKRANGGELKTGYLSIVMDPDELPLDEHCERLPYDASKWEFPRDRLKLGKPLGRGAFGQVIEADAFGIDKTATCRTVAVKMLKEGATHSEHRALMSELKILIHIGHHLNVVNLLGACTKPGGPLMVIVEFCKFGNLSTYLRSKRNEFVPYKTKGARFRQGKDYVGAIPVDLKRRLDSITSSQSSASSGFVEEKSLSDVEEEEAPEDLYKDFLTLEHLICYSFQVAKGMEFLASRKCIHRDLAARNILLSEKNVVKICDFGLARDIYKDPDYVRKGDARLPLKWMAPETIFDRVYTIQSDVWSFGVLLWEIFSLGASPYPGVKIDEEFCRRLKEGTRMRAPDYTTPEMYQTMLDCWHGEPSQRPTFSELVEHLGNLLQANAQQDGKDYIVLPISETLSMEEDSGLSLPTSPVSCMEEEEVCDPKFHYDNTAGISQYLQNSKRKSRPVSVKTFEDIPLEEPEVKVIPDDNQTDSGMVLASEELKTLEDRTKLSPSFGGMVPSKSRESVASEGSNQTSGYQSGYHSDDTDTTVYSSEEAELLKLIEIGVQTGSTAQILQPDSGTTLSSPPV cd44_human CD44 antigen Epican CDw44 Phagocytic glycoprotein I Extracellular matrix receptor III HUTCH-I Phagocytic glycoprotein 1 GP90 lymphocyte homing/adhesion receptor Hermes antigen Hyaluronate receptor Heparan sulfate proteoglycan CD44 LHR MDU2 MDU3 MIC4 protein protein human Homo sapiens Human MDKFWWHAAWGLCLVPLSLAQIDLNITCRFAGVFHVEKNGRYSISRTEAADLCKAFNSTLPTMAQMEKALSIGFETCRYGFIEGHVVIPRIHPNSICAANNTGVYILTSNTSQYDTYCFNASAPPEEDCTSVTDLPNAFDGPITITIVNRDGTRYVQKGEYRTNPEDIYPSNPTDDDVSSGSSSERSSTSGGYIFYTFSTVHPIPDEDSPWITDSTDRIPATTLMSTSATATETATKRQETWDWFSWLFLPSESKNHLHTTTQMAGTSSNTISAGWEPNEENEDERDRHLSFSGSGIDDDEDFISSTISTTPRAFDHTKQNQDWTQWNPSHSNPEVLLQTTTRMTDVDRNGTTAYEGNWNPEAHPPLIHHEHHEEEETPHSTSTIQATPSSTTEETATQKEQWFGNRWHEGYRQTPKEDSHSTTGTAAASAHTSHPMQGRTTPSPEDSSWTDFFNPISHPMGRGHQAGRRMDMDSSHSITLQPTANPNTGLVEDLDRTGPLSMTTQQSNSQSFSTSHEGLEEDKDHPTTSTLTSSNRNDVTGGRRDPNHSEGSTTLLEGYTSHYPHTKESRTFIPVTSAKTGSFGVTAVTVGDSNSNVNRSLSGDQDTFHPSGGSHTTHGSESDGHSHGSQEGGANTTSGPIRTPQIPEWLIILASLLALALILAVCIAVNSRRRCGQKKKLVINSGNGAVEDRKPSGLNGEASKSQEMVHLVNKESSETPDQFMTADETRNLQNVDMKIGV hgf_human Hepatocyte growth factor HGF HPTA Scatter factor Hepatopoietin-A protein protein human Homo sapiens Human MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPALKIKTKKVNTADQCANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGHEFDLYENKDYIRNCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGKDLQENYCRNPRGEEGGPWCFTSNPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTESGKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAIKTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPENFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNYMGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGNPLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRYRNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQLVYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDGLLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKMRMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS hgf-5 1314923 n/a 1314930 unspecified role unspecified role neutral component neutral component P14210-6:p.[Lys60Glu;Lys62Glu;Arg73Glu] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig 3A lanes 4+8 18 kDa heparin or 4.2 kDa Heparan Sulphate (HS) oligomers supported NK1 WT multimer formation, but HS tetramers (1.2 kDa) did not. Oligomers were not detected using the mutant HS-NK1 3S. 18 kDa heparin, 4.2 kDa HS. hgf-7 1314923 n/a 1314930 unspecified role unspecified role neutral component neutral component P14210-6:p.[Lys60Glu;Lys62Glu;Arg73Glu] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 3.0 direct interaction direct interaction false false false Fig 3A lanes 4+8 18 kDa heparin or 4.2 kDa Heparan Sulphate (HS) oligomers supported NK1 WT multimer formation, but HS tetramers (1.2 kDa) did not. Oligomers were not detected using the mutant HS-NK1 3S. 18 kDa heparin, 4.2 kDa HS. hgf-6 1314923 n/a 1314930 unspecified role unspecified role neutral component neutral component P14210-6:p.[Lys60Glu;Lys62Glu;Arg73Glu] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 direct interaction direct interaction false false false Fig 3A lanes 4+8 18 kDa heparin or 4.2 kDa Heparan Sulphate (HS) oligomers supported NK1 WT multimer formation, but HS tetramers (1.2 kDa) did not. Oligomers were not detected using the mutant HS-NK1 3S. 18 kDa heparin, 4.2 kDa HS. nrp1-vegfa-1 1314923 n/a 1314931 unspecified role unspecified role neutral component neutral component P15692-4:p.[Arg149Glu;Arg150Glu;Arg185Glu] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. n/a 1314932 unspecified role unspecified role neutral component neutral component c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false human Homo sapiens Human physical association physical association false false false Fig S5D Multimers (~250kDa) of VEGF/NRP1-Fc were observed for both WT and mutant VEGF. nrp1-vegfa-6 1314923 n/a 1314931 unspecified role unspecified role neutral component neutral component human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. Stoichiometry: 2.0 physical association physical association false false false Fig S5D nrp1-vegfa-3 1314923 n/a 1314931 unspecified role unspecified role neutral component neutral component P15692-4:p.[Arg149Glu;Arg150Glu;Arg185Glu] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. Stoichiometry: 2.0 n/a 1314932 unspecified role unspecified role neutral component neutral component c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false human Homo sapiens Human physical association physical association false false false Fig S5D A 160 kDa band, consistent with a 1:1 complex of NRP1-Fc (~120kDa) and VEGF dimer (~44kDa), was detected. nrp1-vegfa 1314923 n/a 1314932 unspecified role unspecified role neutral component neutral component c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false human Homo sapiens Human n/a 1314931 unspecified role unspecified role neutral component neutral component human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. physical association physical association false false false S5D Multimers(~250kDa) of VEGF/NRP1-Fc were observed for both WT and mutant VEGF. nrp1-vegfa-4 1314923 n/a 1314931 unspecified role unspecified role neutral component neutral component human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. Stoichiometry: 2.0 n/a 1314932 unspecified role unspecified role neutral component neutral component c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false human Homo sapiens Human physical association physical association false false false S5D A 160 kDa band, consistent with a 1:1 complex of NRP1-Fc (~120kDa) and VEGF dimer (~44kDa), was detected. met-hgf 1314924 n/a 1314933 unspecified role unspecified role bait bait ectodomain(extracellular)_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c_terminal_igg1_constant_region(pro100-lys330) fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range false c_terminus 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 false mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc n/a 1314930 unspecified role unspecified role prey prey ruthenium_tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P14210-6:p.[Lys60Glu;Lys62Glu;Arg73Glu] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig 2B Saturation binding of tagged NK1 3S to the Met-Ig protein was unaffected by the absence of HS. met-hgf-2 1314924 n/a 1314930 unspecified role unspecified role prey prey ruthenium_tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1314933 unspecified role unspecified role bait bait c_terminus 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 false ectodomain(extracellular)_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c_terminal_igg1_constant_region(pro100-lys330) fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range false mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc n/a 1314934 unspecified role unspecified role ancillary ancillary physical association physical association false false false Fig 2C NK1 WT binding to Met-Ig in the absence of Heparan Sulphate was significantly compromised. heparin-hgf-3 1314924 n/a 1314930 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P14210-6:p.[Lys60Glu;Lys62Glu;Arg73Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ruthenium_tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1314935 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false pig Sus scrofa (Pig) Pig direct interaction direct interaction false false false Fig 2A vegfa-kdr-2 1314924 n/a 1314931 unspecified role unspecified role bait bait P15692-4:p.[Arg149Glu;Arg150Glu;Arg185Glu] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. Stoichiometry: 2.0 n/a 1314936 unspecified role unspecified role prey prey ectodomain(extracellular)_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c_terminal_igg1_constant_region(pro100-lys330) fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range false c_terminus 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 false mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc physical association physical association false false false Fig 6C vegfa-kdr-1 1314924 n/a 1314936 unspecified role unspecified role prey prey c_terminus 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 false c_terminal_igg1_constant_region(pro100-lys330) fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ectodomain(extracellular)_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc n/a 1314931 unspecified role unspecified role bait bait human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. physical association physical association false false false Fig 6C vegfa_dimer 1314925 n/a 1314931 unspecified role unspecified role neutral component neutral component human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. Stoichiometry: 2.0 physical association physical association false false false Fig S5F Dimers were observed for both WT and mutant VEGF in the presence and absence of Heparin beads. s3_vegfa 1314925 n/a 1314931 unspecified role unspecified role neutral component neutral component P15692-4:p.[Arg149Glu;Arg150Glu;Arg185Glu] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. Stoichiometry: 2.0 physical association physical association false false false Fig S5F-2 Dimers were observed for both WT and mutant VEGF in the presence and absence of Heparin beads. heparin-vegfa 1314925 n/a 1314931 unspecified role unspecified role neutral component neutral component human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. n/a 1314935 unspecified role unspecified role ancillary ancillary pig Sus scrofa (Pig) Pig n/a 1314931 unspecified role unspecified role neutral component neutral component human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. physical association physical association false false false Fig S5F Multimers were observed only for WT VEGF in the presence of Heparin beads. cd44-met-1 1314926 n/a 1314930 unspecified role unspecified role ancillary ancillary P14210-6:p.[Lys60Glu;Lys62Glu;Arg73Glu] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 3.0 n/a 1314937 unspecified role unspecified role bait bait n/a 1314933 unspecified role unspecified role prey prey physical association physical association false false false Fig 4C cd44-met-2 1314926 n/a 1314933 unspecified role unspecified role prey prey n/a 1314937 unspecified role unspecified role bait bait n/a 1314938 unspecified role unspecified role ancillary ancillary physical association physical association false false false Fig 4C cd44-met-4 1314927 n/a 1314937 unspecified role unspecified role bait bait n/a 1314930 competitor competitor competitor ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P14210-6:p.[Lys60Glu;Lys62Glu;Arg73Glu] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 3.0 n/a 1314933 unspecified role unspecified role prey prey n/a 1314938 unspecified role unspecified role ancillary ancillary physical association physical association false false false Fig 4D vegf 1314928 n/a 1314931 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. Stoichiometry: 2.0 direct interaction direct interaction false false false Fig 6B - right image heparin-hgf-4 1314929 n/a 1314935 unspecified role unspecified role bait bait n/a 1314930 unspecified role unspecified role prey prey P14210-6:p.[Lys60Glu;Lys62Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P14210-6:p.Arg73Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P14210-6:p.[Lys60Glu;Lys62Glu;Arg73Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Table S2