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 Tyrosine phosphorylation of the orphan receptor ESDN/DCBLD2 serves as a scaffold for the signaling adaptor CrkL. Tyrosine phosphorylation of the orphan receptor ESDN/DCBLD2 serves as a scaffold for the signaling adaptor CrkL. FEBS Lett. (0014-5793) 2013 Aten TM., Redmond MM., Weaver SO., Love CC., Joy RM., Lapp AS., Rivera OD., Hinkle KL., Ballif BA. in vitro In vitro pull down pull down experimental particp experimental participant identification experimental particp Aten TM., Redmond MM., Weaver SO., Love CC., Joy RM., Lapp AS., Rivera OD., Hinkle KL., Ballif BA. FEBS Lett. (0014-5793) 2013 fyn_human Tyrosine-protein kinase Fyn p59-Fyn Proto-oncogene c-Fyn Src-like kinase FYN Proto-oncogene Syn protein protein human Homo sapiens Human MGCVQCKDKEATKLTEERDGSLNQSSGYRYGTDPTPQHYPSFGVTSIPNYNNFHAAGGQGLTVFGGVNSSSHTGTLRTRGGTGVTLFVALYDYEARTEDDLSFHKGEKFQILNSSEGDWWEARSLTTGETGYIPSNYVAPVDSIQAEEWYFGKLGRKDAERQLLSFGNPRGTFLIRESETTKGAYSLSIRDWDDMKGDHVKHYKIRKLDNGGYYITTRAQFETLQQLVQHYSERAAGLCCRLVVPCHKGMPRLTDLSVKTKDVWEIPRESLQLIKRLGNGQFGEVWMGTWNGNTKVAIKTLKPGTMSPESFLEEAQIMKKLKHDKLVQLYAVVSEEPIYIVTEYMNKGSLLDFLKDGEGRALKLPNLVDMAAQVAAGMAYIERMNYIHRDLRSANILVGNGLICKIADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELVTKGRVPYPGMNNREVLEQVERGYRMPCPQDCPISLHELMIHCWKKDPEERPTFEYLQSFLEDYFTATEPQYQPGENL DIP protein P06241 original sequence version: 195 DIP protein P06241 original sequence version: 147 DIP protein P06241 original sequence version: 164 4A1E443A4B5A0977 crkl_human Crk-like protein CRKL protein protein human Homo sapiens Human MSSARFDSSDRSAWYMGPVSRQEAQTRLQGQRHGMFLVRDSSTCPGDYVLSVSENSRVSHYIINSLPNRRFKIGDQEFDHLPALLEFYKIHYLDTTTLIEPAPRYPSPPMGSVSAPNLPTAEDNLEYVRTLYDFPGNDAEDLPFKKGEILVIIEKPEEQWWSARNKDGRVGMIPVPYVEKLVRSSPHGKHGNRNSNSYGIPEPAHAYAQPQTTTPLPAVSGSPGAAITPLPSTQNGPVFAKAIQKRVPCAYDKTALALEVGDIVKVTRMNINGQWEGEVNGRKGLFPFTHVKIFDPQNPDENE DIP protein P46109 original sequence version: 64 DIP protein P46109 original sequence version: 103 DIP protein P46109 original sequence version: 104 DIP protein P46109 original sequence version: 156 294CF1EE2CD44B81 dcbd2_human Discoidin, CUB and LCCL domain-containing protein 2 DCBLD2 CLCP1 CUB, LCCL and coagulation factor V/VIII-homology domains protein 1 ESDN Endothelial and smooth muscle cell-derived neuropilin-like protein protein protein human Homo sapiens Human MASRAVVRARRCPQCPQVRAAAAAPAWAALPLSRSLPPCSNSSSFSMPLFLLLLLVLLLLLEDAGAQQGDGCGHTVLGPESGTLTSINYPQTYPNSTVCEWEIRVKMGERVRIKFGDFDIEDSDSCHFNYLRIYNGIGVSRTEIGKYCGLGLQMNHSIESKGNEITLLFMSGIHVSGRGFLASYSVIDKQDLITCLDTASNFLEPEFSKYCPAGCLLPFAEISGTIPHGYRDSSPLCMAGVHAGVVSNTLGGQISVVISKGIPYYESSLANNVTSVVGHLSTSLFTFKTSGCYGTLGMESGVIADPQITASSVLEWTDHTGQENSWKPKKARLKKPGPPWAAFATDEYQWLQIDLNKEKKITGIITTGSTMVEHNYYVSAYRILYSDDGQKWTVYREPGVEQDKIFQGNKDYHQDVRNNFLPPIIARFIRVNPTQWQQKIAMKMELLGCQFIPKGRPPKLTQPPPPRNSNDLKNTTAPPKIAKGRAPKFTQPLQPRSSNEFPAQTEQTTASPDIRNTTVTPNVTKDVALAAVLVPVLVMVLTTLILILVCAWHWRNRKKKTEGTYDLPYWDRAGWWKGMKQFLPAKAVDHEETPVRYSSSEVNHLSPREVTTVLQADSAEYAQPLVGGIVGTLHQRSTFKPEEGKEAGYADLDPYNSPGQEVYHAYAEPLPITGPEYATPIIMDMSGHPTTSVGQPSTSTFKATGNQPPPLVGTYNTLLSRTDSCSSAQAQYDTPKAGKPGLPAPDELVYQVPQSTQEVSGAGRDGECDVFKEIL mint homomint 3D06F81EF2337010 bcar1_human Breast cancer anti-estrogen resistance protein 1 CRK-associated substrate p130cas BCAR1 CAS CRKAS CASS1 Cas scaffolding protein family member 1 protein protein human Homo sapiens Human MNHLNVLAKALYDNVAESPDELSFRKGDIMTVLEQDTQGLDGWWLCSLHGRQGIVPGNRLKILVGMYDKKPAGPGPGPPATPAQPQPGLHAPAPPASQYTPMLPNTYQPQPDSVYLVPTPSKAQQGLYQVPGPSPQFQSPPAKQTSTFSKQTPHHPFPSPATDLYQVPPGPGGPAQDIYQVPPSAGMGHDIYQVPPSMDTRSWEGTKPPAKVVVPTRVGQGYVYEAAQPEQDEYDIPRHLLAPGPQDIYDVPPVRGLLPSQYGQEVYDTPPMAVKGPNGRDPLLEVYDVPPSVEKGLPPSNHHAVYDVPPSVSKDVPDGPLLREETYDVPPAFAKAKPFDPARTPLVLAAPPPDSPPAEDVYDVPPPAPDLYDVPPGLRRPGPGTLYDVPRERVLPPEVADGGVVDSGVYAVPPPAEREAPAEGKRLSASSTGSTRSSQSASSLEVAGPGREPLELEVAVEALARLQQGVSATVAHLLDLAGSAGATGSWRSPSEPQEPLVQDLQAAVAAVQSAVHELLEFARSAVGNAAHTSDRALHAKLSRQLQKMEDVHQTLVAHGQALDAGRGGSGATLEDLDRLVACSRAVPEDAKQLASFLHGNASLLFRRTKATAPGPEGGGTLHPNPTDKTSSIQSRPLPSPPKFTSQDSPDGQYENSEGGWMEDYDYVHLQGKEEFEKTQKELLEKGSITRQGKSQLELQQLKQFERLEQEVSRPIDHDLANWTPAQPLAPGRTGGLGPSDRQLLLFYLEQCEANLTTLTNAVDAFFTAVATNQPPKIFVAHSKFVILSAHKLVFIGDTLSRQAKAADVRSQVTHYSNLLCDLLRGIVATTKAAALQYPSPSAAQDMVERVKELGHSTQQFRRVLGQLAAA DIP protein P56945 original sequence version: 113 DIP protein P56945 original sequence version: 116 B81EC3430049E795 crkl-bcar1 1 ESDN 4 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level CAS 5 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level CrkL-SH2 3 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst 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 association association f1 homomint domino mint SILAC ratios were determined for all peptides from proteins uniquely bound to the CrkL-SH2 domain. Figure 1B shows the two proteins that had SILAC ratios that were statistically different from those of all other identified proteins. These two proteins, Cas and ESDN, presented a profile consistent with SFK-dependent CrkL-SH2 binding crkl-dcbld2 1 ESDN 4 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate Q96PD2:p.Tyr621Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:58:29 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 17:37:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr621phe' Tue Jun 30 20:17:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr621Phe' Q96PD2:p.Tyr750Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:23:10 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 15:45:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr750phe' Tue Jun 30 18:04:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr750Phe' Q96PD2:p.Tyr666Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:23:04 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 14:48:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr666phe' Tue Jun 30 16:29:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr666Phe' Q96PD2:p.Tyr677Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:58:27 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 20:10:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr677phe' Tue Jun 30 20:24:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr677Phe' Q96PD2:p.Tyr565Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:58:24 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 16:24:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr565phe' Tue Jun 30 16:40:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr565Phe' flag tag 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 Q96PD2:p.Tyr655Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:23:02 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 19:44:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr655phe' Tue Jun 30 16:38:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr655Phe' Q96PD2:p.Tyr732Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:22:56 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 16:57:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr732phe' Tue Jun 30 16:19:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr732Phe' CrkL-SH2 3 weight by staining molecular weight estimation by staining weight by staining experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association homomint domino mint To determine if ESDN underwent reversible tyrosine phosphorylation at YxxP tyrosines we immunoprecipitated ESDN-Flag from the same extracts used in the GST-CrkL-SH2 pulldowns (Fig. 2C). All constructs, including the Y7F, showed reversible tyrosine phosphorylation as measured by anti-phosphotyrosine immunblotting with the Y7F showing dramatically reduced anti-phosphotyrosine immunoreactivity (Fig. 2C). Pulldown assays with both GST-CrkL-SH2 and GST-Fyn-SH2 showed H2O2-induced binding to wild type ESDN but not to ESDN Y7F (Fig. 3A). Importantly, similar results were obtained when ESDN was co-expressed with Fyn in the absence of H2O2 Fig. 3B) f2c f3a f3b f4c fyn-dcbld2 1 ESDN 4 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate Q96PD2:p.Tyr666Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:20:11 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 19:48:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr666phe' Tue Jun 30 19:31:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr666Phe' Q96PD2:p.Tyr732Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:20:11 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 18:54:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr732phe' Tue Jun 30 17:42:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr732Phe' Q96PD2:p.Tyr677Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:56:52 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 20:01:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr677phe' Tue Jun 30 16:46:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr677Phe' Q96PD2:p.Tyr750Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:56:48 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 14:46:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr750phe' Tue Jun 30 19:36:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr750Phe' Q96PD2:p.Tyr655Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:56:49 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 18:56:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr655phe' Tue Jun 30 19:46:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr655Phe' Q96PD2:p.Tyr621Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:57:07 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 14:48:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr621phe' Tue Jun 30 17:04:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr621Phe' Q96PD2:p.Tyr565Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Tue Jul 12 10:20:35 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 17:46:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr565phe' Tue Jun 30 17:51:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr565Phe' flag tag 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 Fyn-SH2 2 weight by staining molecular weight estimation by staining weight by staining experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association homomint domino mint f3a f3b Pulldown assays with both GST-CrkL-SH2 and GST-Fyn-SH2 showed H2O2-induced binding to wild type ESDN but not to ESDN Y7F (Fig. 3A). Importantly, similar results were obtained when ESDN was co-expressed with Fyn in the absence of H2O2 Fig. 3B) crkl-dcbld2-1 1 ESDN 4 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate flag tag 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 CrkL-SH2 3 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst 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 physical association physical association f3c homomint domino mint However, while pharmacological inhibition of SFKs with the highly selective Src Inhibitor-1 (Src-1) [19] led to increased ESDN electrophoretic mobility, significant binding of ESDN to the CrkL-SH2 domain was retained (Fig. 3C), suggesting that additional kinases are important in promoting the ESDN-CrkL interaction.