bhf-ucl Gremlin is a novel agonist of the major proangiogenic receptor VEGFR2. Gremlin is a novel agonist of the major proangiogenic receptor VEGFR2. Blood (0006-4971) 2010 imex curation Mitola S., Ravelli C., Moroni E., Salvi V., Leali D., Ballmer-Hofer K., Zammataro L., Presta M. Cardiac - Interactions involving cardiac related proteins presta@med.unibs.it 18-Feb-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa predetermined predetermined participant predetermined Mitola S., Ravelli C., Moroni E., Salvi V., Leali D., Ballmer-Hofer K., Zammataro L., Presta M. Cardiac - Interactions involving cardiac related proteins Plates coated with sVEGFR1-Fc or sVEGFR2-Fc were incubated with VEGF-A in presence of different competitors. The nature of the participants (human proteins) was confirmed by the authors, except for VEGF-E. Therefore its interaction as a competitor was not annotated. presta@med.unibs.it Accepted 2014-FEB-12 by ORCHARD Blood (0006-4971) 2010 imex curation Gremlin is a novel agonist of the major proangiogenic receptor VEGFR2. Gremlin is a novel agonist of the major proangiogenic receptor VEGFR2. Blood (0006-4971) 2010 imex curation Mitola S., Ravelli C., Moroni E., Salvi V., Leali D., Ballmer-Hofer K., Zammataro L., Presta M. Cardiac - Interactions involving cardiac related proteins presta@med.unibs.it 18-Feb-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Mitola S., Ravelli C., Moroni E., Salvi V., Leali D., Ballmer-Hofer K., Zammataro L., Presta M. Cardiac - Interactions involving cardiac related proteins Gremlin was injected over the sVEGFR2-coated sensor chip in the absence or in the presence of different competitors. Origin of proteins taken from manufacturers websites with additional information from subsequent papers by the author PMID:21921258 presta@med.unibs.it Accepted 2014-FEB-12 by ORCHARD Blood (0006-4971) 2010 imex curation Gremlin is a novel agonist of the major proangiogenic receptor VEGFR2. Gremlin is a novel agonist of the major proangiogenic receptor VEGFR2. Blood (0006-4971) 2010 imex curation Mitola S., Ravelli C., Moroni E., Salvi V., Leali D., Ballmer-Hofer K., Zammataro L., Presta M. Cardiac - Interactions involving cardiac related proteins presta@med.unibs.it 18-Feb-2014: Contacted by IntAct-Help. Only protein-protein interactions human-uvec Human Umbilical Vein Endothelial Cells (HUVEC) huvec umbilical_vein_endothelium Umbilical vein endothelial cell [Umbilical vein endothelium] crosslink cross-linking study crosslink tag perox activity tag visualisation by peroxidase activity tag perox activity Mitola S., Ravelli C., Moroni E., Salvi V., Leali D., Ballmer-Hofer K., Zammataro L., Presta M. Cardiac - Interactions involving cardiac related proteins HUVECs were incubated with gremlin conjugated with the bifunctional photoactivable biotin-label transfer cross-linker Sulfo-SBED Biotin Label transfer reagent (in absence or in the presence of unlabeled gremlin or VEFG-A). The cell lysates were immunoprecipitated with anti-VEGFR2 antibody. Immunoprecipitated samples were separated by SDS electrophoresis and probed with streptavidin-horseradish peroxidase to visualize the biotin-VEGFR2 complex. presta@med.unibs.it Accepted 2014-FEB-12 by ORCHARD Blood (0006-4971) 2010 imex curation vgfr1_human Vascular endothelial growth factor receptor 1 Vascular permeability factor receptor Tyrosine-protein kinase receptor FLT Tyrosine-protein kinase FRT Fms-like tyrosine kinase 1 FLT1 FLT FRT VEGFR1 protein protein human Homo sapiens Human MVSYWDTGVLLCALLSCLLLTGSSSGSKLKDPELSLKGTQHIMQAGQTLHLQCRGEAAHKWSLPEMVSKESERLSITKSACGRNGKQFCSTLTLNTAQANHTGFYSCKYLAVPTSKKKETESAIYIFISDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVQISTPRPVKLLRGHTLVLNCTATTPLNTRVQMTWSYPDEKNKRASVRRRIDQSNSHANIFYSVLTIDKMQNKDKGLYTCRVRSGPSFKSVNTSVHIYDKAFITVKHRKQQVLETVAGKRSYRLSMKVKAFPSPEVVWLKDGLPATEKSARYLTRGYSLIIKDVTEEDAGNYTILLSIKQSNVFKNLTATLIVNVKPQIYEKAVSSFPDPALYPLGSRQILTCTAYGIPQPTIKWFWHPCNHNHSEARCDFCSNNEESFILDADSNMGNRIESITQRMAIIEGKNKMASTLVVADSRISGIYICIASNKVGTVGRNISFYITDVPNGFHVNLEKMPTEGEDLKLSCTVNKFLYRDVTWILLRTVNNRTMHYSISKQKMAITKEHSITLNLTIMNVSLQDSGTYACRARNVYTGEEILQKKEITIRDQEAPYLLRNLSDHTVAISSSTTLDCHANGVPEPQITWFKNNHKIQQEPGIILGPGSSTLFIERVTEEDEGVYHCKATNQKGSVESSAYLTVQGTSDKSNLELITLTCTCVAATLFWLLLTLFIRKMKRSSSEIKTDYLSIIMDPDEVPLDEQCERLPYDASKWEFARERLKLGKSLGRGAFGKVVQASAFGIKKSPTCRTVAVKMLKEGATASEYKALMTELKILTHIGHHLNVVNLLGACTKQGGPLMVIVEYCKYGNLSNYLKSKRDLFFLNKDAALHMEPKKEKMEPGLEQGKKPRLDSVTSSESFASSGFQEDKSLSDVEEEEDSDGFYKEPITMEDLISYSFQVARGMEFLSSRKCIHRDLAARNILLSENNVVKICDFGLARDIYKNPDYVRKGDTRLPLKWMAPESIFDKIYSTKSDVWSYGVLLWEIFSLGGSPYPGVQMDEDFCSRLREGMRMRAPEYSTPEIYQIMLDCWHRDPKERPRFAELVEKLGDLLQANVQQDGKDYIPINAILTGNSGFTYSTPAFSEDFFKESISAPKFNSGSSDDVRYVNAFKFMSLERIKTFEELLPNATSMFDDYQGDSSTLLASPMLKRFTWTDSKPKASLKIDLRVTSKSKESGLSDVSRPSFCHSSCGHVSEGKRRFTYDHAELERKIACCSPPPDYNSVVLYSTPPI FF3381EEFAF0787C 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 grem1_human Gremlin-1 GREM1 CKTSF1B1 DAND2 DRM PIG2 Cysteine knot superfamily 1, BMP antagonist 1 Increased in high glucose protein 2 Down-regulated in Mos-transformed cells protein DAN domain family member 2 Cell proliferation-inducing gene 2 protein protein protein human Homo sapiens Human MSRTAYTVGALLLLLGTLLPAAEGKKKGSQGAIPPPDKAQHNDSEQTQSPQQPGSRNRGRGQGRGTAMPGEEVLESSQEALHVTERKYLKRDWCKTQPLKQTIHEEGCNSRTIINRFCYGQCNSFYIPRHIRKEEGSFQSCSFCKPKKFTTMMVTLNCPELQPPTKKKRVTRVKQCRCISIDLD 4B588598DE12C47E 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 plgf_human Placenta growth factor PGF PLGF PGFL protein protein human Homo sapiens Human MPVMRLFPCFLQLLAGLALPAVPPQQWALSAGNGSSEVEVVPFQEVWGRSYCRALERLVDVVSEYPSEVEHMFSPSCVSLLRCTGCCGDENLHCVPVETANVTMQLLKIRSGDRPSYVELTFSQHVRCECRHSPGRQSPDMPGDFRADAPSFLPPRRSLPMLFRMEWGCALTGSQSAVWPSSPVPEEIPRMHPGRNGKKQQRKPLREKMKPERCGDAVPRR D364C6A73C1C6987 kdr-grem1-2 2 5 competitor competitor competitor prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain fc fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm physical association physical association Fig.1B flt1-vegfa-14 1 7 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain fc fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined sVEGFR1fc (P17948) used in this experiment is a commercial one from RELIATech GmbH. It has an extra aminoacid (Arg) between Thr751 and Ser752 direct interaction direct interaction Fig.1C grem1-vgfr2 3 6 unspecified role unspecified role neutral component neutral component biotin biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role neutral component neutral component human-uvec Human Umbilical Vein Endothelial Cells (HUVEC) huvec umbilical_vein_endothelium Umbilical vein endothelial cell [Umbilical vein endothelium] physical association physical association fig.1D The VEGFR2 biotinylation by sulfo-gremlin was inhibited by an excess of unlabeled gremlin ( O60565) or VEGF-A (P15692) that were acting as antagonists. kdr-grem1-3 2 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction Fig.1A O60565 increases activity of P35968 kdr-vegfa-13 1 5 unspecified role unspecified role bait bait fc fc-igg tag fc-igg 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 7 unspecified role unspecified role prey prey direct interaction direct interaction Fig.1 C P15692 increases activity of P35968 kdr-vegfa-14 1 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain fc fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 competitor competitor competitor prey prey physical association physical association Fig.1 C flt1-pgf-2 1 7 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait fc fc-igg tag fc-igg 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 sVEGFR1fc (P17948) used in this experiment is a commercial one from RELIATech GmbH. It has an extra aminoacid (Arg) between Thr751 and Ser752 8 competitor competitor competitor prey prey ecoli Escherichia coli (strain K12) physical association physical association Fig.1C