IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Dissociation of SHP-1 from spinophilin during platelet activation exposes an inhibitory binding site for protein phosphatase-1 (PP1). Dissociation of SHP-1 from spinophilin during platelet activation exposes an inhibitory binding site for protein phosphatase-1 (PP1). PloS one 2014 imex curation Ma P., Foote DC., Sinnamon AJ., Brass LF. Results from figs 2A, 4, 5, 6A were not annotated due to lack of antibody specificity (PP1). some human - but trasfected SPL is RAT. peisong@mail.med.upenn.edu,brass@mail.med.upenn.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-patu Human-PATU Pancreatic Adenocarcinoma cell line patu PATU-Human Pancreatic adenocarcinoma cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ma P., Foote DC., Sinnamon AJ., Brass LF. Results from figs 2A, 4, 5, 6A were not annotated due to lack of antibody specificity (PP1). some human - but trasfected SPL is RAT. peisong@mail.med.upenn.edu,brass@mail.med.upenn.edu Accepted 2018-JUN-26 AT 13:24 BST AT 13:24 BST by PPORRAS PloS one 2014 Only protein-protein interactions imex curation imex curation added; I have added more interactions for Fig 3B: cannot add PP1 - in 3B LHS image- as is endogenous, and no idea which one. The SHP1/SPL interaction is in the next experiment. I cannot see any interaction in which SPL is a competitor.; Figure 6C is phosphorylation - not an IP. -as before, I cannot add PP1 as a competitor here (fig 3B) as it is endogenous, and identity unclear. Dissociation of SHP-1 from spinophilin during platelet activation exposes an inhibitory binding site for protein phosphatase-1 (PP1). Dissociation of SHP-1 from spinophilin during platelet activation exposes an inhibitory binding site for protein phosphatase-1 (PP1). PloS one 2014 imex curation Ma P., Foote DC., Sinnamon AJ., Brass LF. Results from figs 2A, 4, 5, 6A were not annotated due to lack of antibody specificity (PP1). some human - but trasfected SPL is RAT. peisong@mail.med.upenn.edu,brass@mail.med.upenn.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Ma P., Foote DC., Sinnamon AJ., Brass LF. Results from figs 2A, 4, 5, 6A were not annotated due to lack of antibody specificity (PP1). some human - but trasfected SPL is RAT. peisong@mail.med.upenn.edu,brass@mail.med.upenn.edu Accepted 2018-JUN-26 AT 13:24 BST AT 13:24 BST by PPORRAS PloS one 2014 Only protein-protein interactions imex curation imex curation added; I have added more interactions for Fig 3B: cannot add PP1 - in 3B LHS image- as is endogenous, and no idea which one. The SHP1/SPL interaction is in the next experiment. I cannot see any interaction in which SPL is a competitor.; Figure 6C is phosphorylation - not an IP. -as before, I cannot add PP1 as a competitor here (fig 3B) as it is endogenous, and identity unclear. Dissociation of SHP-1 from spinophilin during platelet activation exposes an inhibitory binding site for protein phosphatase-1 (PP1). Dissociation of SHP-1 from spinophilin during platelet activation exposes an inhibitory binding site for protein phosphatase-1 (PP1). PloS one 2014 imex curation Ma P., Foote DC., Sinnamon AJ., Brass LF. Results from figs 2A, 4, 5, 6A were not annotated due to lack of antibody specificity (PP1). some human - but trasfected SPL is RAT. peisong@mail.med.upenn.edu,brass@mail.med.upenn.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Ma P., Foote DC., Sinnamon AJ., Brass LF. Results from figs 2A, 4, 5, 6A were not annotated due to lack of antibody specificity (PP1). some human - but trasfected SPL is RAT. peisong@mail.med.upenn.edu,brass@mail.med.upenn.edu Accepted 2018-JUN-26 AT 13:24 BST AT 13:24 BST by PPORRAS PloS one 2014 Only protein-protein interactions imex curation imex curation added; I have added more interactions for Fig 3B: cannot add PP1 - in 3B LHS image- as is endogenous, and no idea which one. The SHP1/SPL interaction is in the next experiment. I cannot see any interaction in which SPL is a competitor.; Figure 6C is phosphorylation - not an IP. -as before, I cannot add PP1 as a competitor here (fig 3B) as it is endogenous, and identity unclear. neb2_rat Neurabin-2 Ppp1r9b Neurabin-II Neural tissue-specific F-actin-binding protein II p130 PP1bp134 Protein phosphatase 1 regulatory subunit 9B Spinophilin Spino protein protein rat Rattus norvegicus (Rat) Rat MMKTEPRGPGGPLRSASPHRSAYEAGIQALKPPDAPGPDEAPKAAHHKKYGSNVHRIKSMFLQMGTTTGPPGEAGGASGMAEAPRASDRGVRLSLPRASSLNENVDHSALLKLGTSVSERVSRFDSKPAPSAQPAPPPHPPSRLQETRKLFERSVPAASGGDKEAVARRLLRQERASLQDRKLDVVVRFNGSTEALDKLDADAVSPTVSQLSAVFEKADSRTGLHRAPGPPRAAGAPQVNSKLVTKRSRVFQPPPPPPAPSGDAATEKDRGPGGQQPPQHRVAPARPPPKPREVRKIKPVEVEESGESEAESAPGEVIQAEVTVHAALENGSTTATTASPAPEEPKAEAVPEEEASSSVATLERGVDNGRAPDMAPEEVDESKKEDFSEADLVDVSAYSGLGEDSGGSALEEDDEEDEEDGEPPYEPESGCVEIPGLSEEEDPAPSRKIHFSTAPIQVFSTYSNEDYDRRNEDVDPMAASAEYELEKRVERLELFPVELEKDSEGLGISIIGMGAGADMGLEKLGIFVKTVTEGGAAHRDGRIQVNDLLVEVDGTSLVGVTQSFAASVLRNTKGRVRFMIGRERPGEQSEVAQLIQQTLEQERWQREMMEQRYAQYGEDDEETGEYATDEDEELSPTFPGGEMAIEVFELAENEDALSPVEMEPEKLVHKFKELQIKHAVTEAEIQQLKRKLQSLEQEKGRWRVEKAQLEQSVEENKERMEKLEGYWGEAQSLCQAVDEHLRETQAQYQALERKYSKAKRLIKDYQQKEIEFLKKETAQRRVLEESELARKEEMDKLLDKISELEGNLQTLRNSNST DIP protein O35274 original sequence version: 87 D31D3E94EE2FFB6A pp1b_human Serine/threonine-protein phosphatase PP1-beta catalytic subunit PPP1CB protein protein human Homo sapiens Human MADGELNVDSLITRLLEVRGCRPGKIVQMTEAEVRGLCIKSREIFLSQPILLELEAPLKICGDIHGQYTDLLRLFEYGGFPPEANYLFLGDYVDRGKQSLETICLLLAYKIKYPENFFLLRGNHECASINRIYGFYDECKRRFNIKLWKTFTDCFNCLPIAAIVDEKIFCCHGGLSPDLQSMEQIRRIMRPTDVPDTGLLCDLLWSDPDKDVQGWGENDRGVSFTFGADVVSKFLNRHDLDLICRAHQVVEDGYEFFAKRQLVTLFSAPNYCGEFDNAGGMMSVDETLMCSFQILKPSEKKAKYQYGGLNSGRPVTPPRTANPPKKR DIP protein P62140 original sequence version: 130 E8356022E9B94ECD ptn6_human Tyrosine-protein phosphatase non-receptor type 6 PTPN6 HCP Protein-tyrosine phosphatase 1C Hematopoietic cell protein-tyrosine phosphatase SH-PTP1 Protein-tyrosine phosphatase SHP-1 PTP1C protein protein human Homo sapiens Human MVRWFHRDLSGLDAETLLKGRGVHGSFLARPSRKNQGDFSLSVRVGDQVTHIRIQNSGDFYDLYGGEKFATLTELVEYYTQQQGVLQDRDGTIIHLKYPLNCSDPTSERWYHGHMSGGQAETLLQAKGEPWTFLVRESLSQPGDFVLSVLSDQPKAGPGSPLRVTHIKVMCEGGRYTVGGLETFDSLTDLVEHFKKTGIEEASGAFVYLRQPYYATRVNAADIENRVLELNKKQESEDTAKAGFWEEFESLQKQEVKNLHQRLEGQRPENKGKNRYKNILPFDHSRVILQGRDSNIPGSDYINANYIKNQLLGPDENAKTYIASQGCLEATVNDFWQMAWQENSRVIVMTTREVEKGRNKCVPYWPEVGMQRAYGPYSVTNCGEHDTTEYKLRTLQVSPLDNGDLIREIWHYQYLSWPDHGVPSEPGGVLSFLDQINQRQESLPHAGPIIVHCSAGIGRTGTIIVIDMLMENISTKGLDCDIDIQKTIQMVRAQRSGMVQTEAQYKFIYVAIAQFIETTKKKLEVLQSQKGQESEYGNITYPPAMKNAHAKASRTSSKHKEDVYENLHTKNKREEKVKKQRSADKEKSKGSLKRK DIP protein P29350 original sequence version: 183 DIP protein P29350 original sequence version: 139 DIP protein P29350-1 original sequence version: 139 4D7736C21D3542D2 pp1a_human Serine/threonine-protein phosphatase PP1-alpha catalytic subunit PPP1CA PPP1A protein protein human Homo sapiens Human MSDSEKLNLDSIIGRLLEVQGSRPGKNVQLTENEIRGLCLKSREIFLSQPILLELEAPLKICGDIHGQYYDLLRLFEYGGFPPESNYLFLGDYVDRGKQSLETICLLLAYKIKYPENFFLLRGNHECASINRIYGFYDECKRRYNIKLWKTFTDCFNCLPIAAIVDEKIFCCHGGLSPDLQSMEQIRRIMRPTDVPDQGLLCDLLWSDPDKDVQGWGENDRGVSFTFGAEVVAKFLHKHDLDLICRAHQVVEDGYEFFAKRQLVTLFSAPNYCGEFDNAGAMMSVDETLMCSFQILKPADKNKGKYGQFSGLNPGGRPITPPRNSAKAKK DIP protein P62136 original sequence version: 72 DIP protein P62136 original sequence version: 76 DIP protein P62136 original sequence version: 123 DIP protein P62136 original sequence version: 126 DIP protein P62136-1 original sequence version: 98 60C37E1AD9831DAC pp1g_human Serine/threonine-protein phosphatase PP1-gamma catalytic subunit PPP1CC Protein phosphatase 1C catalytic subunit protein protein human Homo sapiens Human MADLDKLNIDSIIQRLLEVRGSKPGKNVQLQENEIRGLCLKSREIFLSQPILLELEAPLKICGDIHGQYYDLLRLFEYGGFPPESNYLFLGDYVDRGKQSLETICLLLAYKIKYPENFFLLRGNHECASINRIYGFYDECKRRYNIKLWKTFTDCFNCLPIAAIVDEKIFCCHGGLSPDLQSMEQIRRIMRPTDVPDQGLLCDLLWSDPDKDVLGWGENDRGVSFTFGAEVVAKFLHKHDLDLICRAHQVVEDGYEFFAKRQLVTLFSAPNYCGEFDNAGAMMSVDETLMCSFQILKPAEKKKPNATRPVTPPRGMITKQAKK DIP protein P36873 original sequence version: 169 DIP protein P36873 original sequence version: 177 0EEEF0E842188536 neb2_human Neurabin-2 Neurabin-II Spinophilin Protein phosphatase 1 regulatory subunit 9B PPP1R9B PPP1R6 protein protein human Homo sapiens Human MMKTEPRGPGGPLRSASPHRSAYEAGIQALKPPDAPGPDEAPKGAHHKKYGSNVHRIKSMFLQMGTTAGPSGEAGGGAGLAEAPRASERGVRLSLPRASSLNENVDHSALLKLGTSVSERVSRFDSKPAPSAQPAPPPHPPSRLQETRKLFERSAPAAAGGDKEAAARRLLRQERAGLQDRKLDVVVRFNGSTEALDKLDADAVSPTVSQLSAVFEKADSRTGLHRGPGLPRAAGVPQVNSKLVSKRSRVFQPPPPPPPAPSGDAPAEKERCPAGQQPPQHRVAPARPPPKPREVRKIKPVEVEESGESEAESAPGEVIQAEVTVHAALENGSTVATAASPAPEEPKAQAAPEKEAAAVAPPERGVGNGRAPDVAPEEVDESKKEDFSEADLVDVSAYSGLGEDSAGSALEEDDEDDEEDGEPPYEPESGCVEIPGLSEEEDPAPSRKIHFSTAPIQVFSTYSNEDYDRRNEDVDPMAASAEYELEKRVERLELFPVELEKDSEGLGISIIGMGAGADMGLEKLGIFVKTVTEGGAAHRDGRIQVNDLLVEVDGTSLVGVTQSFAASVLRNTKGRVRFMIGRERPGEQSEVAQLIQQTLEQERWQREMMEQRYAQYGEDDEETGEYATDEDEELSPTFPGGEMAIEVFELAENEDALSPVDMEPEKLVHKFKELQIKHAVTEAEIQQLKRKLQSLEQEKGRWRVEKAQLEQSVEENKERMEKLEGYWGEAQSLCQAVDEHLRETQAQYQALERKYSKAKRLIKDYQQKEIEFLKKETAQRRVLEESELARKEEMDKLLDKISELEGNLQTLRNSNST 706A359FE8382DBB 4567667ityu-1 1 9 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role bait bait cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Figure 1 PAR1 agonist peptide; SFLLRN Adding the PAR1 (thrombin receptor) agonist peptide, SFLLRN, caused an approximately 10-fold increase in binding. Adding the PAR1 (thrombin receptor) agonist peptide, SFLLRN, caused an approximately 10-fold increase in binding ppp1ca-ppp1r9b-7 1 9 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role bait bait cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Figure 1-2 PAR1 agonist peptide; SFLLRN Adding the PAR1 (thrombin receptor) agonist peptide, SFLLRN, caused an approximately 10-fold increase in binding. ppp1ca-ppp1r9b-8 2 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate region 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 Figure 2B, Supp fig 1 ppp1cc-ppp1r9b-3 2 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate region 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 Figure 2B ppp1ca-ppp1r9b-9 2 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate O35274:p.Phe451Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain F A Wed May 02 15:21:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe451ala' Tue Jun 30 16:10:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe451Ala' region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region 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 Figure 3B, 6B In CHO cells transfected with spinophilin and SHP-1 (but not PP1), SHP-1 binding to spinophilin was favored over endogenous PP1. However, when PP1a was over-expressed, it displaced SHP-1. In CHO cells transfected with spinophilin and SHP-1 (but not PP1), SHP-1 binding to spinophilin was favored over endogenous PP1 (Fig. 3B, left lane in the gel stack shown). However, when PP1a was over-expressed, it displaced SHP-1 ppp1r9b-ppp1ca-1 3 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 3B-2 In CHO cells transfected with spinophilin and SHP-1 (but not PP1), SHP-1 binding to spinophilin was favored over endogenous PP1. However, when PP1a was over-expressed, it displaced SHP-1. ppp1cc-ppp1r9b-4 1 5 unspecified role unspecified role bait bait cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Figure 1-3 PAR1 peptide; SFLLRN ppp1r9b-ppp1ca-2 2 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 competitor competitor competitor prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region 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 association association Figure 3Bi In CHO cells transfected with spinophilin and SHP-1 (but not PP1), SHP-1 binding to spinophilin was favored over endogenous PP1. However, when PP1a was over-expressed, it displaced SHP-1. In CHO cells transfected with spinophilin and SHP-1 (but not PP1), SHP-1 binding to spinophilin was favored over endogenous PP1 (Fig. 3B, left lane in the gel stack shown). However, when PP1a was over-expressed, it displaced SHP-1