MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. EMBO J. (0261-4189) 2006 imex curation Shintani Y., Takashima S., Asano Y., Kato H., Liao Y., Yamazaki S., Tsukamoto O., Seguchi O., Yamamoto H., Fukushima T., Sugahara K., Kitakaze M., Hori M. 2009-01-08 Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Shintani Y., Takashima S., Asano Y., Kato H., Liao Y., Yamazaki S., Tsukamoto O., Seguchi O., Yamamoto H., Fukushima T., Sugahara K., Kitakaze M., Hori M. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. EMBO J. (0261-4189) 2006 imex curation Shintani Y., Takashima S., Asano Y., Kato H., Liao Y., Yamazaki S., Tsukamoto O., Seguchi O., Yamamoto H., Fukushima T., Sugahara K., Kitakaze M., Hori M. 2009-01-08 Only protein-protein interactions in vitro In vitro crosslink cross-linking study crosslink nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Shintani Y., Takashima S., Asano Y., Kato H., Liao Y., Yamazaki S., Tsukamoto O., Seguchi O., Yamamoto H., Fukushima T., Sugahara K., Kitakaze M., Hori M. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. EMBO J. (0261-4189) 2006 imex curation Shintani Y., Takashima S., Asano Y., Kato H., Liao Y., Yamazaki S., Tsukamoto O., Seguchi O., Yamamoto H., Fukushima T., Sugahara K., Kitakaze M., Hori M. 2009-01-08 Only protein-protein interactions human-smooth muscle Homo sapiens smooth_muscle smooth muscle anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Shintani Y., Takashima S., Asano Y., Kato H., Liao Y., Yamazaki S., Tsukamoto O., Seguchi O., Yamamoto H., Fukushima T., Sugahara K., Kitakaze M., Hori M. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. Glycosaminoglycan modification of neuropilin-1 modulates VEGFR2 signaling. EMBO J. (0261-4189) 2006 imex curation Shintani Y., Takashima S., Asano Y., Kato H., Liao Y., Yamazaki S., Tsukamoto O., Seguchi O., Yamamoto H., Fukushima T., Sugahara K., Kitakaze M., Hori M. 2009-01-08 Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Shintani Y., Takashima S., Asano Y., Kato H., Liao Y., Yamazaki S., Tsukamoto O., Seguchi O., Yamamoto H., Fukushima T., Sugahara K., Kitakaze M., Hori M. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation 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 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 kdr-vegfa-2 2 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 1.0 physical association physical association F1A EMBO-Journal When human coronary artery smooth muscle cells (CASMCs) were incubated with 125I-labeled VEGF, we detected two distinct binding proteins after cell-surface crosslinking (Figure 1A). homomint mint nrp1-vegfa-7 2 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 1.0 physical association physical association F1A EMBO-Journal When human coronary artery smooth muscle cells (CASMCs) were incubated with 125I-labeled VEGF, we detected two distinct binding proteins after cell-surface crosslinking (Figure 1A). homomint mint nrp1-vegfa-8 4 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate flag tagged flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F3B EMBO-Journal we measured the binding of 125I-labeled VEGF to FLAG-tagged NRP1 in these cells. After incubation with 125I-labeled VEGF, cell lysates were immunoprecipitated with an anti-FLAG antibody, and bound radioactivity was counted. NRP1 WT\' bound VEGF with 3.87- and 2.27-fold higher than NRP1 S612A in SMCs and ECs, respectively (Figure 3B). homomint mint nrp1-vegfa-9 1 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 physical association physical association F2E EMBO-Journal We confirmed that addition of FLAG tag to NRP1 does not affect VEGF binding (data not shown) and that the mutation itself (NRP1 S612A) did not change VEGF binding to the core protein of NRP1 (Figure 2E). homomint mint kdr-nrp1-1 3 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell v5 tagged v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4E EMBO-Journal We next examined the ability of GAG-modified NRP1 to associate with VEGFR2 in the presence of VEGF by co-immunoprecipitation. After pretreatment with heparitinase and/or chondroitinase, V5-tagged VEGFR2 was precipitated from SMC lysates in the presence of VEGF. As shown in Figure 4E, CS-modified NRP1 minimally associated with VEGFR2 compared to non-modified or HS-modified NRP1. homomint mint