bhf-ucl mitola-2010-2 Gremlin is a novel agonist of the major proangiogenic receptor VEGFR2. 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 Blood (0006-4971) 2010 imex curation Only protein-protein interactions mitola-2010-1 Gremlin is a novel agonist of the major proangiogenic receptor VEGFR2. 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 Blood (0006-4971) 2010 imex curation Only protein-protein interactions mitola-2010-3 Gremlin is a novel agonist of the major proangiogenic receptor VEGFR2. 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 Blood (0006-4971) 2010 imex curation Only protein-protein interactions 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 vegfa_human Vascular endothelial growth factor A Vascular permeability factor VEGFA VEGF protein protein human Homo sapiens Human MNFLLSWVHWSLALLLYLHHAKWSQAAPMAEGGGQNHHEVVKFMDVYQRSYCHPIETLVDIFQEYPDEIEYIFKPSCVPLMRCGGCCNDEGLECVPTEESNITMQIMRIKPHQGQHIGEMSFLQHNKCECRPKKDRARQEKKSVRGKGKGQKRKRKKSRYKSWSVYVGARCCLMPWSLPGPHPCGPCSERRKHLFVQDPQTCKCSCKNTDSRCKARQLELNERTCRCDKPRR 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 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 plgf_human Placenta growth factor PGF PLGF PGFL protein protein human Homo sapiens Human MPVMRLFPCFLQLLAGLALPAVPPQQWALSAGNGSSEVEVVPFQEVWGRSYCRALERLVDVVSEYPSEVEHMFSPSCVSLLRCTGCCGDENLHCVPVETANVTMQLLKIRSGDRPSYVELTFSQHVRCECRHSPGRQSPDMPGDFRADAPSFLPPRRSLPMLFRMEWGCALTGSQSAVWPSSPVPEEIPRMHPGRNGKKQQRKPLREKMKPERCGDAVPRR kdr-vegfa-13 1477331 n/a 1477334 unspecified role unspecified role bait bait fc fc-igg tag fc-igg 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 n/a 1477335 unspecified role unspecified role prey prey direct interaction direct interaction false false false Fig.1 C P15692 increases activity of P35968 kdr-vegfa-14 1477331 n/a 1477335 unspecified role unspecified role prey prey n/a 1477334 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 false fc fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1477336 competitor competitor competitor prey prey physical association physical association false false false Fig.1 C flt1-vegfa-14 1477331 n/a 1477335 unspecified role unspecified role prey prey n/a 1477337 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 false fc fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 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 false false false Fig.1C flt1-pgf-2 1477331 n/a 1477335 unspecified role unspecified role prey prey n/a 1477337 unspecified role unspecified role bait bait fc fc-igg tag fc-igg 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 sVEGFR1fc (P17948) used in this experiment is a commercial one from RELIATech GmbH. It has an extra aminoacid (Arg) between Thr751 and Ser752 n/a 1477338 competitor competitor competitor prey prey ecoli Escherichia coli (strain K12) physical association physical association false false false Fig.1C kdr-grem1-2 1477332 n/a 1477334 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 false fc fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1477336 unspecified role unspecified role prey prey n/a 1477334 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 false spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm physical association physical association false false false Fig.1B kdr-grem1-3 1477332 n/a 1477336 unspecified role unspecified role prey prey n/a 1477334 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 false spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction false false false 1477332 Fig.1A O60565 increases activity of P35968 grem1-vgfr2 1477333 n/a 1477336 unspecified role unspecified role neutral component neutral component biotin biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1477334 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 false false false 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.