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/ Blocking neuropilin-1 function has an additive effect with anti-VEGF to inhibit tumor growth. Blocking neuropilin-1 function has an additive effect with anti-VEGF to inhibit tumor growth. Cancer Cell (1535-6108) 2007 imex curation Pan Q., Chanthery Y., Liang WC., Stawicki S., Mak J., Rathore N., Tong RK., Kowalski J., Yee SF., Pacheco G., Ross S., Cheng Z., Le Couter J., Plowman G., Peale F., Koch AW., Wu Y., Bagri A., Tessier-Lavigne M., Watts RJ. Cancer - Interactions investigated in the context of cancer To find the range for the VEGF165 construct the reference Jia et al 2006 was consulted. rwatts@gene.com 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 2 human-uvec Human Umbilical Vein Endothelial Cells (HUVEC) huvec umbilical_vein_endothelium Umbilical vein endothelial cell [Umbilical vein endothelium] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Pan Q., Chanthery Y., Liang WC., Stawicki S., Mak J., Rathore N., Tong RK., Kowalski J., Yee SF., Pacheco G., Ross S., Cheng Z., Le Couter J., Plowman G., Peale F., Koch AW., Wu Y., Bagri A., Tessier-Lavigne M., Watts RJ. Cancer - Interactions investigated in the context of cancer To find the range for the VEGF165 construct the reference Jia et al 2006 was consulted. rwatts@gene.com Accepted 2007-FEB-15 by JYOTI. 02-MAR-2006: Contacted by JYOTI. Cancer Cell (1535-6108) 2007 Only protein-protein interactions 2 imex curation Blocking neuropilin-1 function has an additive effect with anti-VEGF to inhibit tumor growth. Blocking neuropilin-1 function has an additive effect with anti-VEGF to inhibit tumor growth. Cancer Cell (1535-6108) 2007 imex curation Pan Q., Chanthery Y., Liang WC., Stawicki S., Mak J., Rathore N., Tong RK., Kowalski J., Yee SF., Pacheco G., Ross S., Cheng Z., Le Couter J., Plowman G., Peale F., Koch AW., Wu Y., Bagri A., Tessier-Lavigne M., Watts RJ. Cancer - Interactions investigated in the context of cancer To find the range for the VEGF165 construct the reference Jia et al 2006 was consulted. rwatts@gene.com 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 2 in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Pan Q., Chanthery Y., Liang WC., Stawicki S., Mak J., Rathore N., Tong RK., Kowalski J., Yee SF., Pacheco G., Ross S., Cheng Z., Le Couter J., Plowman G., Peale F., Koch AW., Wu Y., Bagri A., Tessier-Lavigne M., Watts RJ. Cancer - Interactions investigated in the context of cancer To find the range for the VEGF165 construct the reference Jia et al 2006 was consulted. rwatts@gene.com Accepted 2007-FEB-15 by JYOTI. 02-MAR-2006: Contacted by JYOTI. Cancer Cell (1535-6108) 2007 Only protein-protein interactions 2 imex curation vegfa_human Vascular endothelial growth factor A Vascular permeability factor VEGFA VEGF protein protein human Homo sapiens Human MNFLLSWVHWSLALLLYLHHAKWSQAAPMAEGGGQNHHEVVKFMDVYQRSYCHPIETLVDIFQEYPDEIEYIFKPSCVPLMRCGGCCNDEGLECVPTEESNITMQIMRIKPHQGQHIGEMSFLQHNKCECRPKKDRARQEKKSVRGKGKGQKRKRKKSRYKSWSVYVGARCCLMPWSLPGPHPCGPCSERRKHLFVQDPQTCKCSCKNTDSRCKARQLELNERTCRCDKPRR DIP protein P15692 original sequence version: 190 DIP protein P15692 original sequence version: 150 FB49F364446F4D01 nrp1_human Neuropilin-1 Vascular endothelial cell growth factor 165 receptor NRP1 NRP VEGF165R protein protein human Homo sapiens Human MERGLPLLCAVLALVLAPAGAFRNDKCGDTIKIESPGYLTSPGYPHSYHPSEKCEWLIQAPDPYQRIMINFNPHFDLEDRDCKYDYVEVFDGENENGHFRGKFCGKIAPPPVVSSGPFLFIKFVSDYETHGAGFSIRYEIFKRGPECSQNYTTPSGVIKSPGFPEKYPNSLECTYIVFVPKMSEIILEFESFDLEPDSNPPGGMFCRYDRLEIWDGFPDVGPHIGRYCGQKTPGRIRSSSGILSMVFYTDSAIAKEGFSANYSVLQSSVSEDFKCMEALGMESGEIHSDQITASSQYSTNWSAERSRLNYPENGWTPGEDSYREWIQVDLGLLRFVTAVGTQGAISKETKKKYYVKTYKIDVSSNGEDWITIKEGNKPVLFQGNTNPTDVVVAVFPKPLITRFVRIKPATWETGISMRFEVYGCKITDYPCSGMLGMVSGLISDSQITSSNQGDRNWMPENIRLVTSRSGWALPPAPHSYINEWLQIDLGEEKIVRGIIIQGGKHRENKVFMRKFKIGYSNNGSDWKMIMDDSKRKAKSFEGNNNYDTPELRTFPALSTRFIRIYPERATHGGLGLRMELLGCEVEAPTAGPTTPNGNLVDECDDDQANCHSGTGDDFQLTGGTTVLATEKPTVIDSTIQSEFPTYGFNCEFGWGSHKTFCHWEHDNHVQLKWSVLTSKTGPIQDHTGDGNFIYSQADENQKGKVARLVSPVVYSQNSAHCMTFWYHMSGSHVGTLRVKLRYQKPEEYDQLVWMAIGHQGDHWKEGRVLLHKSLKLYQVIFEGEIGKGNLGGIAVDDISINNHISQEDCAKPADLDKKNPEIKIDETGSTPGYEGEGEGDKNISRKPGNVLKTLDPILITIIAMSALGVLLGAVCGVVLYCACWHNGMSERNLSALENYNFELVDGVKLKKDKLNTQSTYSEA DIP protein O14786 original sequence version: 114 1EAC2FA6C8FD6A0B 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 DIP protein P35968 original sequence version: 81 DIP protein P35968 original sequence version: 146 DIP protein P35968 original sequence version: 117 DIP protein P35968 original sequence version: 121 59E7C44B05CFEBB3 nrp1-vegf 2 3 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 4 unspecified role unspecified role bait bait fc Human NRP1 fused to human Fc fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 1 C Anti-NRP1B strongly blocked VEGF binding to NRP1 and ECs, whereas anti-NRP1A did not This interaction was inhibited by the anti-NRPB antibody (which targets the primary region for VEGF binding - the coagulation V/VIII factor (or b1-b2) domains). The anti-NRPA antibody (which targets the primary region for Sem3A binding - the CUB (or a1-a2) domains) does not inhibit the interaction. kdr-nrp1 1 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association 4 F in the presence of VEGF, NRP1 can be coimmunoprecipitated from HUVEC lysate with anti- VEGFR2 antibodies (Figure 4F, lane 2). This interaction was reduced by anti-VEGF (Figure 4F, lane 3). Interestingly, both anti-NRP1A and anti-NRP1B strongly inhibit the formation of the NRP1/VEGFR2 complex This interaction took place in the presence of VEGF and was reduced by anti-VEGF. This interaction was reduced by anti-VEGF. The interaction was also strongly inhibted by both anti-NRP1A (targets the primary Sem3A binding region) and anti-NRP1B antibodies (targets the primary VEGF binding region).