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 Insight into the Sts-1 - mediated regulation of TCR signaling through the use of novel substrate-trapping Sts-1 phosphatase variants. Insight into the Sts-1 - mediated regulation of TCR signaling through the use of novel substrate-trapping Sts-1 phosphatase variants. FEBS J. (1742-464X) 2013 imex curation Luis B.S., Carpino N. nicholas.carpino@stonybrook.edu 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Luis B.S., Carpino N. nicholas.carpino@stonybrook.edu Accepted 2013-NOV-11 by LUANA FEBS J. (1742-464X) 2013 Only protein-protein interactions imex curation Insight into the Sts-1 - mediated regulation of TCR signaling through the use of novel substrate-trapping Sts-1 phosphatase variants. Insight into the Sts-1 - mediated regulation of TCR signaling through the use of novel substrate-trapping Sts-1 phosphatase variants. FEBS J. (1742-464X) 2013 imex curation Luis B.S., Carpino N. nicholas.carpino@stonybrook.edu 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Luis B.S., Carpino N. nicholas.carpino@stonybrook.edu Accepted 2013-NOV-11 by LUANA FEBS J. (1742-464X) 2013 Only protein-protein interactions imex curation Insight into the Sts-1 - mediated regulation of TCR signaling through the use of novel substrate-trapping Sts-1 phosphatase variants. Insight into the Sts-1 - mediated regulation of TCR signaling through the use of novel substrate-trapping Sts-1 phosphatase variants. FEBS J. (1742-464X) 2013 imex curation Luis B.S., Carpino N. nicholas.carpino@stonybrook.edu 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Luis B.S., Carpino N. nicholas.carpino@stonybrook.edu Accepted 2013-NOV-11 by LUANA FEBS J. (1742-464X) 2013 Only protein-protein interactions imex curation Insight into the Sts-1 - mediated regulation of TCR signaling through the use of novel substrate-trapping Sts-1 phosphatase variants. Insight into the Sts-1 - mediated regulation of TCR signaling through the use of novel substrate-trapping Sts-1 phosphatase variants. FEBS J. (1742-464X) 2013 imex curation Luis B.S., Carpino N. nicholas.carpino@stonybrook.edu 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Luis B.S., Carpino N. nicholas.carpino@stonybrook.edu Accepted 2013-NOV-11 by LUANA FEBS J. (1742-464X) 2013 Only protein-protein interactions imex curation ubs3b_mouse Ubiquitin-associated and SH3 domain-containing protein B Ubash3b Sts1 Cbl-interacting protein p70 Suppressor of T-cell receptor signaling 1 T-cell ubiquitin ligand 2 Tyrosine-protein phosphatase STS1/TULA2 protein protein mouse Mus musculus Mouse MAAREELYSKVTPRRDRLQRPGTVKHGSALDVLLSMGFPRARAQKALASTGGRSVQAACDWLFSHVGDPFLDDPLPREYVLYLRPTGPLAQKLSDFWQQSKQICGKNKAHNIFPHITLCQFFMCEDSKVDALGEALQTTVSRWKCKFSAPLPLELYTSSNFIGLFVKEDSAEVLKKFAADFAAEAASKTEVHVEPHKKQLHVTLAYHFQASHLPTLEKLAQNIDVKLGCDWVATIFSRDIRFANHETLQVIYPYSPQNDDELELVPGDFIFMSPMEQTSTSEGWIYGTSLTTGCSGLLPENYITKADECSTWIFHGSYSILNTVSSSSLAFGDGALERRQYEDQGLGETTPLTIICQPMQPLRVNSQPGPQKRCLFVCRHGERMDVVFGKYWLSQCFDAKGRYIRTNLNMPHSLPQRSGGFRDYEKDAPITVFGCMQARLVGEALLESNTVIDHVYCSPSLRCVQTAHNILKGLQQDNHLKIRVEPGLFEWTKWVAGSTLPAWIPPSELAAANLSVDTTYRPHIPVSKLAISESYDTYINRSFQVTKEIISECKSKGNNILIVAHASSLEACTCQLQGLSPQNSKDFVQMVRKIPYLGFCSCEELGETGIWQLTDPPILPLTHGPTGGFNWRETLLQE 9B665F45AF208EF6 zap70_human Tyrosine-protein kinase ZAP-70 ZAP70 SRK Syk-related tyrosine kinase 70 kDa zeta-chain associated protein protein protein human Homo sapiens Human MPDPAAHLPFFYGSISRAEAEEHLKLAGMADGLFLLRQCLRSLGGYVLSLVHDVRFHHFPIERQLNGTYAIAGGKAHCGPAELCEFYSRDPDGLPCNLRKPCNRPSGLEPQPGVFDCLRDAMVRDYVRQTWKLEGEALEQAIISQAPQVEKLIATTAHERMPWYHSSLTREEAERKLYSGAQTDGKFLLRPRKEQGTYALSLIYGKTVYHYLISQDKAGKYCIPEGTKFDTLWQLVEYLKLKADGLIYCLKEACPNSSASNASGAAAPTLPAHPSTLTHPQRRIDTLNSDGYTPEPARITSPDKPRPMPMDTSVYESPYSDPEELKDKKLFLKRDNLLIADIELGCGNFGSVRQGVYRMRKKQIDVAIKVLKQGTEKADTEEMMREAQIMHQLDNPYIVRLIGVCQAEALMLVMEMAGGGPLHKFLVGKREEIPVSNVAELLHQVSMGMKYLEEKNFVHRDLAARNVLLVNRHYAKISDFGLSKALGADDSYYTARSAGKWPLKWYAPECINFRKFSSRSDVWSYGVTMWEALSYGQKPYKKMKGPEVMAFIEQGKRMECPPECPPELYALMSDCWIYKWEDRPDFLTVEQRMRACYYSLASKVEGPPGSTQKAEAACA DIP protein P43403 original sequence version: 146 DIP protein P43403-1 original sequence version: 167 D1E1A8EC66FA116F ubash3b-zap70 1 Sts-1 5 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his380cys_arg462ala_glu490gln trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain Zap-70 6 predetermined predetermined participant predetermined anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ptyr319 porphres porphyrin modified residue PorphRes certain certain sequence position certain certain certain sequence position certain physical association physical association f1c We began by assessing the interaction between a Zap-70-derived phosphopeptide (GSVYESPpYSDPEEL) and the different Sts-1PGM mutants described above. Pulldown assays were performed in which phosphopeptide-coupled beads were added to lysates prepared from cells expressing wild-type or mutant Sts-1PGM, and bound proteins were assessed by immunoblot analysis. We observed that four Sts-1PGM mutants interacted stably with the bound phosphopeptide, asevidenced by their ability to be isolated from the lysate (Figure 1C). Interestingly, the four mutants appeared to interact with the phospho-peptide beads with substantially different affinities. Specifically, Sts-1PGM H380C single mutant exhibited very weak interaction with the phospho-peptide beads, while the double mutants Sts-1PGM H380C/E490A and Sts-1PGM H380C/E490Q appeared to have the strongest affinity and the Sts-1PGM H380C/R462A/E490Q triple mutant was precipitated to an intermediate degree. Full-length Sts-1 variants also displayed similar binding characteristics (data not shown), with double mutants H380C/E490A and H380C/E490Q (henceforth referred to as CA and CQ respectively) displaying increased binding capabilities than the single mutant H380C (henceforth referred to as C). These results suggest that the CA and CQ mutants could serve as significantly better substrate-trapping enzymes than the C mutant. zap70-ubash3b 1 Sts-1 5 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate his380cys_glu490ala trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 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 Zap-70 6 predetermined predetermined participant predetermined anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ptyr319 porphres porphyrin modified residue PorphRes certain certain sequence position certain certain certain sequence position certain physical association physical association f1c We began by assessing the interaction between a Zap-70-derived phosphopeptide (GSVYESPpYSDPEEL) and the different Sts-1PGM mutants described above. Pulldown assays were performed in which phosphopeptide-coupled beads were added to lysates prepared from cells expressing wild-type or mutant Sts-1PGM, and bound proteins were assessed by immunoblot analysis. We observed that four Sts-1PGM mutants interacted stably with the bound phosphopeptide, asevidenced by their ability to be isolated from the lysate (Figure 1C). Interestingly, the four mutants appeared to interact with the phospho-peptide beads with substantially different affinities. Specifically, Sts-1PGM H380C single mutant exhibited very weak interaction with the phospho-peptide beads, while the double mutants Sts-1PGM H380C/E490A and Sts-1PGM H380C/E490Q appeared to have the strongest affinity and the Sts-1PGM H380C/R462A/E490Q triple mutant was precipitated to an intermediate degree. Full-length Sts-1 variants also displayed similar binding characteristics (data not shown), with double mutants H380C/E490A and H380C/E490Q (henceforth referred to as CA and CQ respectively) displaying increased binding capabilities than the single mutant H380C (henceforth referred to as C). These results suggest that the CA and CQ mutants could serve as significantly better substrate-trapping enzymes than the C mutant. zap70-ubash3b-1 1 Zap-70 6 predetermined predetermined participant predetermined anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ptyr319 porphres porphyrin modified residue PorphRes certain certain sequence position certain certain certain sequence position certain Sts-1 5 anti tag western anti tag western blot anti tag western anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his380cys_glu490gln trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 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 physical association physical association f1c We began by assessing the interaction between a Zap-70-derived phosphopeptide (GSVYESPpYSDPEEL) and the different Sts-1PGM mutants described above. Pulldown assays were performed in which phosphopeptide-coupled beads were added to lysates prepared from cells expressing wild-type or mutant Sts-1PGM, and bound proteins were assessed by immunoblot analysis. We observed that four Sts-1PGM mutants interacted stably with the bound phosphopeptide, asevidenced by their ability to be isolated from the lysate (Figure 1C). Interestingly, the four mutants appeared to interact with the phospho-peptide beads with substantially different affinities. Specifically, Sts-1PGM H380C single mutant exhibited very weak interaction with the phospho-peptide beads, while the double mutants Sts-1PGM H380C/E490A and Sts-1PGM H380C/E490Q appeared to have the strongest affinity and the Sts-1PGM H380C/R462A/E490Q triple mutant was precipitated to an intermediate degree. Full-length Sts-1 variants also displayed similar binding characteristics (data not shown), with double mutants H380C/E490A and H380C/E490Q (henceforth referred to as CA and CQ respectively) displaying increased binding capabilities than the single mutant H380C (henceforth referred to as C). These results suggest that the CA and CQ mutants could serve as significantly better substrate-trapping enzymes than the C mutant. ubash3b-zap70-2 4 Zap-70 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell Sts-1 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his380cys_glu490ala trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain physical association physical association f3c ubash3b-zap70-3 4 Zap-70 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell Sts-1 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his380cys_glu490gln trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain physical association physical association f3c ubash3b-zap70-5 2 Sts-1 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain his380cys_glu490gln trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Zap-70 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell physical association physical association f3d zap70-ubash3b-4 2 Zap-70 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 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 his380cys_glu490ala trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Zap-70 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell physical association physical association f3c zap70-ubash3b-3 3 Zap-70 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level ptyr319 phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain Sts-1 5 predetermined predetermined participant predetermined western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait purified purified his380cys__glu490gln trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f4a f4b f4c ubash3b-zap70-6 3 Zap-70 6 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level ptyr319 phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain Sts-1 5 predetermined predetermined participant predetermined western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait purified purified his380cys_glu490ala trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f4b f4c ubash3b-zap70-7 4 Sts-1 5 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell his380cys_glu490gln trap-mutant trapping mutant trap-mutant certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 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 Zap-70 6 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell P43403:p.Tyr319Phe mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 18:43:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr319phe' Tue Jun 30 18:27:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr319Phe' t7 tag t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f4d