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 Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. EMBO Rep. (1469-221X) 2006 imex curation Tsuboi S., Nonoyama S., Ochs HD. 2009-01-08 Only protein-protein interactions 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. anti tag coip anti tag coimmunoprecipitation anti tag coip nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Tsuboi S., Nonoyama S., Ochs HD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. EMBO Rep. (1469-221X) 2006 imex curation Tsuboi S., Nonoyama S., Ochs HD. 2009-01-08 Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Tsuboi S., Nonoyama S., Ochs HD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. EMBO Rep. (1469-221X) 2006 imex curation Tsuboi S., Nonoyama S., Ochs HD. 2009-01-08 Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Tsuboi S., Nonoyama S., Ochs HD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. EMBO Rep. (1469-221X) 2006 imex curation Tsuboi S., Nonoyama S., Ochs HD. 2009-01-08 Only protein-protein interactions human-platelet Homo sapiens platelet, thrombocytes platelet Platelet, thrombocyte anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Tsuboi S., Nonoyama S., Ochs HD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. Wiskott-Aldrich syndrome protein is involved in alphaIIb beta3-mediated cell adhesion. EMBO Rep. (1469-221X) 2006 imex curation Tsuboi S., Nonoyama S., Ochs HD. 2009-01-08 Only protein-protein interactions human-meg-01 Homo sapiens meg-01 meg-01 anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Tsuboi S., Nonoyama S., Ochs HD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation wipf1_human WAS/WASL-interacting protein family member 1 WIPF1 WIP Wiskott-Aldrich syndrome protein-interacting protein Protein PRPL-2 WASPIP protein protein human Homo sapiens Human MPVPPPPAPPPPPTFALANTEKPTLNKTEQAGRNALLSDISKGKKLKKTVTNDRSAPILDKPKGAGAGGGGGGFGGGGGFGGGGGGGGGGSFGGGGPPGLGGLFQAGMPKLRSTANRDNDSGGSRPPLLPPGGRSTSAKPFSPPSGPGRFPVPSPGHRSGPPEPQRNRMPPPRPDVGSKPDSIPPPVPSTPRPIQSSPHNRGSPPVPGGPRQPSPGPTPPPFPGNRGTALGGGSIRQSPLSSSSPFSNRPPLPPTPSRALDDKPPPPPPPVGNRPSIHREAVPPPPPQNNKPPVPSTPRPSASSQAPPPPPPPSRPGPPPLPPSSSGNDETPRLPQRNLSLSSSTPPLPSPGRSGPLPPPPSERPPPPVRDPPGRSGPLPPPPPVSRNGSTSRALPATPQLPSRSGVDSPRSGPRPPLPPDRPSAGAPPPPPPSTSIRNGFQDSPCEDEWESRFYFHPISDLPPPEPYVQTTKSYPSKLARNESRSGSNRRERGAPPLPPIPR DIP protein O43516 original sequence version: 128 DIP protein O43516 original sequence version: 97 DIP protein O43516 original sequence version: 131 09B7A95E6808A46D wasl_human Neural Wiskott-Aldrich syndrome protein WASL protein protein human Homo sapiens Human MSSVQQQPPPPRRVTNVGSLLLTPQENESLFTFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQVALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDPPNGPNLPMATVDIKNPEITTNRFYGPQVNNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELKNLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPPHNSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPSVAVPPPPPNRMYPPPPPALPSSAPSGPPPPPPSVLGVGPVAPPPPPPPPPPPGPPPPPGLPSDGDHQVPTTAGNKAALLDQIREGAQLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVADGQESTPPTPAPTSGIVGALMEVMQKRSKAIHSSDEDEDEDDEEDFEDDDEWED DIP protein O00401 original sequence version: 55 DIP protein O00401 original sequence version: 147 DIP protein O00401 original sequence version: 150 DIP protein O00401 original sequence version: 136 DIP protein O00401 original sequence version: 138 DIP protein O00401 original sequence version: 125 DIP protein O00401 original sequence version: 112 DIP protein O00401 original sequence version: 126 B5895C6055E23049 cib1_human Calcium and integrin-binding protein 1 Calmyrin DNA-PKcs-interacting protein Kinase-interacting protein SNK-interacting protein 2-28 CIB1 CIB KIP PRKDCIP Calcium- and integrin-binding protein protein protein human Homo sapiens Human MGGSGSRLSKELLAEYQDLTFLTKQEILLAHRRFCELLPQEQRSVESSLRAQVPFEQILSLPELKANPFKERICRVFSTSPAKDSLSFEDFLDLLSVFSDTATPDIKSHYAFRIFDFDDDGTLNREDLSRLVNCLTGEGEDTRLSASEMKQLIDNILEESDIDRDGTINLSEFQHVISRSPDFASSFKIVL DIP protein Q99828 original sequence version: 104 9AA19A0DE7AA1E05 ita2_human Integrin alpha-2 Platelet membrane glycoprotein Ia Collagen receptor VLA-2 subunit alpha CD49 antigen-like family member B ITGA2 CD49B protein protein human Homo sapiens Human MGPERTGAAPLPLLLVLALSQGILNCCLAYNVGLPEAKIFSGPSSEQFGYAVQQFINPKGNWLLVGSPWSGFPENRMGDVYKCPVDLSTATCEKLNLQTSTSIPNVTEMKTNMSLGLILTRNMGTGGFLTCGPLWAQQCGNQYYTTGVCSDISPDFQLSASFSPATQPCPSLIDVVVVCDESNSIYPWDAVKNFLEKFVQGLDIGPTKTQVGLIQYANNPRVVFNLNTYKTKEEMIVATSQTSQYGGDLTNTFGAIQYARKYAYSAASGGRRSATKVMVVVTDGESHDGSMLKAVIDQCNHDNILRFGIAVLGYLNRNALDTKNLIKEIKAIASIPTERYFFNVSDEAALLEKAGTLGEQIFSIEGTVQGGDNFQMEMSQVGFSADYSSQNDILMLGAVGAFGWSGTIVQKTSHGHLIFPKQAFDQILQDRNHSSYLGYSVAAISTGESTHFVAGAPRANYTGQIVLYSVNENGNITVIQAHRGDQIGSYFGSVLCSVDVDKDTITDVLLVGAPMYMSDLKKEEGRVYLFTIKKGILGQHQFLEGPEGIENTRFGSAIAALSDINMDGFNDVIVGSPLENQNSGAVYIYNGHQGTIRTKYSQKILGSDGAFRSHLQYFGRSLDGYGDLNGDSITDVSIGAFGQVVQLWSQSIADVAIEASFTPEKITLVNKNAQIILKLCFSAKFRPTKQNNQVAIVYNITLDADGFSSRVTSRGLFKENNERCLQKNMVVNQAQSCPEHIIYIQEPSDVVNSLDLRVDISLENPGTSPALEAYSETAKVFSIPFHKDCGEDGLCISDLVLDVRQIPAAQEQPFIVSNQNKRLTFSVTLKNKRESAYNTGIVVDFSENLFFASFSLPVDGTEVTCQVAASQKSVACDVGYPALKREQQVTFTINFDFNLQNLQNQASLSFQALSESQEENKADNLVNLKIPLLYDAEIHLTRSTNINFYEISSDGNVPSIVHSFEDVGPKFIFSLKVTTGSVPVSMATVIIHIPQYTKEKNPLMYLTGVQTDKAGDISCNADINPLKIGQTSSSVSFKSENFRHTKELNCRTASCSNVTCWLKDVHMKGEYFVNVTTRIWNGTFASSTFQTVQLTAAAEINTYNPEIYVIEDNTVTIPLMIMKPDEKAEVPTGVIIGSIIAGILLLLALVAILWKLGFFKRKYEKMTKNPDEIDETTELSS DIP protein P17301 original sequence version: 108 DIP protein P17301 original sequence version: 186 7E1B7ED968A94070 wasl-cib1 3 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 7 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 binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain range MINT-1894718: original sequence: 1-105 : MSSVQQQPPPPRRVTNVGSLLLTPQENESLFTFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPR tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F1A EMBO-Reports To determine the role of the WASP N terminus in platelet aggregation, we undertook a yeast two-hybrid screen using the WASP N-terminal domain as bait and isolated a complementary DNA encoding the calcium- and integrin-binding protein (CIB). We found that the binding site for CIB on WASP resided in the WASP N terminus (residues 1 105) and that WASP binding required the CIB N terminus (residues 1 113; Fig 1A) homomint mint cib1-wasl 3 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 8 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 tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain range MINT-1894758: original sequence: 1-113 : MGGSGSRLSKELLAEYQDLTFLTKQEILLAHRRFCELLPQEQRSVESSLRAQVPFEQILSLPELKANPFKERICRVFSTSPAKDSLSFEDFLDLLSVFSDTATPDIKSHYAF Stoichiometry: 1.0 physical association physical association F1A EMBO-Reports To determine the role of the WASP N terminus in platelet aggregation, we undertook a yeast two-hybrid screen using the WASP N-terminal domain as bait and isolated a complementary DNA encoding the calcium- and integrin-binding protein (CIB). We found that the binding site for CIB on WASP resided in the WASP N terminus (residues 1 105) and that WASP binding required the CIB N terminus (residues 1 113; Fig 1A) homomint mint wasl-cib1-1 4 7 monoclonal western monoclonal antibody western blot monoclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 8 polyclonal western polyclonal antibody western blot polyclonal western 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 physical association physical association F1B EMBO-Reports Interaction requires 100uM ADP stimulation We examined whether WASP binds to CIB in platelets CIB was detected only in immunoprecipitates obtained from lysates of stimulated platelets in a time-dependent manner, but not resting platelets homomint mint cib1-wasl-1 4 8 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 7 monoclonal western monoclonal antibody western blot monoclonal western 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 physical association physical association EMBO-Reports Interaction requires 100uM ADP stimulation SF10 WASP was also detected only in anti-CIB immunoprecipitates obtained from lysates of stimulated platelets (supplementary Fig S10 online). homomint mint wasl-wipf1-2 4 6 polyclonal western polyclonal antibody western blot polyclonal western 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 7 monoclonal western monoclonal antibody western blot monoclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association EMBO-Reports SF9B WIP co-immnunoprecipitated with WASP in the lysates from both resting and stimulated platelets (supplementary Fig S9B). homomint mint cib1-wasl-2 2 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain range MINT-1894935: original sequence: 1-105 : MSSVQQQPPPPRRVTNVGSLLLTPQENESLFTFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPR myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain range MINT-1894930: original sequence: 1-113 : MGGSGSRLSKELLAEYQDLTFLTKQEILLAHRRFCELLPQEQRSVESSLRAQVPFEQILSLPELKANPFKERICRVFSTSPAKDSLSFEDFLDLLSVFSDTATPDIKSHYAF 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 direct interaction direct interaction SF1 EMBO-Reports Analysis of WASP CIB binding using the surface plasmon resonance-based biosensor, BIACORE 3000 (BIACORE, Piscataway, NJ, USA), indicated that the WASP N terminus bound directly to the CIB N terminus (supplementary Fig S1 online). homomint mint wasl-cib1-2 5 7 monoclonal western monoclonal antibody western blot monoclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 8 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tagged 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 F2A EMBO-Reports When cells were transfected with CIB, CIB co-immunoprecipitated with WASP (Fig 2A, lane 14), indicating that WASP binds to CIB in transfected cells. homomint mint itga2-cib1 5 8 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell Stoichiometry: 1.0 9 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F2D EMBO-Reports In cells expressing CIB, both CIB and WASP co-immunoprecipitated with aIIb (Fig 2D, lanes 4,6). homomint mint itga2-wasl 5 7 monoclonal western monoclonal antibody western blot monoclonal western 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 9 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F2D EMBO-Reports In cells expressing CIB, both CIB and WASP co-immunoprecipitated with aIIb (Fig 2D, lanes 4,6). homomint mint wasl-cib1-3 1 7 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 myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain range MINT-2748659: original sequence: 1-113 : MGGSGSRLSKELLAEYQDLTFLTKQEILLAHRRFCELLPQEQRSVESSLRAQVPFEQILSLPELKANPFKERICRVFSTSPAKDSLSFEDFLDLLSVFSDTATPDIKSHYAF 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 EMBO-Reports SF9 WASP binding to CIB was blocked by the CIB Nterminal fragment in cells (supplementary Fig S9A, lane 4). homomint mint