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/ Protein-tyrosine kinase and GTPase signals cooperate to phosphorylate and activate Wiskott-Aldrich syndrome protein (WASP)/neuronal WASP. Protein-tyrosine kinase and GTPase signals cooperate to phosphorylate and activate Wiskott-Aldrich syndrome protein (WASP)/neuronal WASP. J. Biol. Chem. (0021-9258) 2006 imex curation Torres E., Rosen MK. BioCreative - Critical Assessment of Information Extraction systems in Biology mrosen@biochem.swmed.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Torres E., Rosen MK. BioCreative - Critical Assessment of Information Extraction systems in Biology mrosen@biochem.swmed.edu Accepted 2006-AUG-29 by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Protein-tyrosine kinase and GTPase signals cooperate to phosphorylate and activate Wiskott-Aldrich syndrome protein (WASP)/neuronal WASP. Protein-tyrosine kinase and GTPase signals cooperate to phosphorylate and activate Wiskott-Aldrich syndrome protein (WASP)/neuronal WASP. J. Biol. Chem. (0021-9258) 2006 imex curation Torres E., Rosen MK. BioCreative - Critical Assessment of Information Extraction systems in Biology mrosen@biochem.swmed.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro phosphatase assay phosphatase assay predetermined predetermined participant predetermined Torres E., Rosen MK. BioCreative - Critical Assessment of Information Extraction systems in Biology Revisit when complex editor in place - Fig 4 concerns interaction with ARP2/complex Additional information on PTPN2 taken from commercial suppliers website mrosen@biochem.swmed.edu Accepted 2006-AUG-29 by JYOTI. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation cdc42_human Cell division control protein 42 homolog CDC42 G25K GTP-binding protein protein protein human Homo sapiens Human MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVMIGGEPYTLGLFDTAGQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHHCPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQKGLKNVFDEAILAALEPPEPKKSRRCVLL 51A437E22A4D8FFF ptn2_human Tyrosine-protein phosphatase non-receptor type 2 T-cell protein-tyrosine phosphatase PTPN2 PTPT protein protein human Homo sapiens Human MPTTIEREFEELDTQRRWQPLYLEIRNESHDYPHRVAKFPENRNRNRYRDVSPYDHSRVKLQNAENDYINASLVDIEEAQRSYILTQGPLPNTCCHFWLMVWQQKTKAVVMLNRIVEKESVKCAQYWPTDDQEMLFKETGFSVKLLSEDVKSYYTVHLLQLENINSGETRTISHFHYTTWPDFGVPESPASFLNFLFKVRESGSLNPDHGPAVIHCSAGIGRSGTFSLVDTCLVLMEKGDDINIKQVLLNMRKYRMGLIQTPDQLRFSYMAIIEGAKCIKGDSSIQKRWKELSKEDLSPAFDHSPNKIMTEKYNGNRIGLEEEKLTGDRCTGLSSKMQDTMEENSESALRKRIREDRKATTAQKVQQMKQRLNENERKRKRWLYWQPILTKMGFMSVILVGAFVGWTLFFQQNAL 0207694A4F058E68 lck_human Tyrosine-protein kinase Lck Protein YT16 Proto-oncogene Lck T cell-specific protein-tyrosine kinase LCK Lymphocyte cell-specific protein-tyrosine kinase p56-LCK Leukocyte C-terminal Src kinase protein protein human Homo sapiens Human MGCGCSSHPEDDWMENIDVCENCHYPIVPLDGKGTLLIRNGSEVRDPLVTYEGSNPPASPLQDNLVIALHSYEPSHDGDLGFEKGEQLRILEQSGEWWKAQSLTTGQEGFIPFNFVAKANSLEPEPWFFKNLSRKDAERQLLAPGNTHGSFLIRESESTAGSFSLSVRDFDQNQGEVVKHYKIRNLDNGGFYISPRITFPGLHELVRHYTNASDGLCTRLSRPCQTQKPQKPWWEDEWEVPRETLKLVERLGAGQFGEVWMGYYNGHTKVAVKSLKQGSMSPDAFLAEANLMKQLQHQRLVRLYAVVTQEPIYIITEYMENGSLVDFLKTPSGIKLTINKLLDMAAQIAEGMAFIEERNYIHRDLRAANILVSDTLSCKIADFGLARLIEDNEYTAREGAKFPIKWTAPEAINYGTFTIKSDVWSFGILLTEIVTHGRIPYPGMTNPEVIQNLERGYRMVRPDNCPEELYQLMRLCWKERPEDRPTFDYLRSVLEDFFTATEGQYQPQP DIP protein P06239 original sequence version: 164 DIP protein P06239-1 original sequence version: 193 44BFF0D43FFB420D 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 wasl-lck-1 1 5 enzyme enzyme unspecified role unspecified role spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 enzyme target enzyme target substrate substrate unspecified role unspecified role gbd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Km = 425 uM Kcat = 51 s-1 Fig. 2A, Table 1 Phosphorylation of Tyr-256 of WASL by LCK We measured the rate of phosphate incorporation from ATP by the Lck kinase domain into a series of N-WASP constructs. wasl-lck-2 1 3 unspecified role unspecified role ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 enzyme enzyme unspecified role unspecified role spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 enzyme target enzyme target substrate substrate unspecified role unspecified role vca domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig. 2B, Table 1 Presence of CDC42 increased efficiency of phosphorylation of N-WASP by LCK by 40-fold. Phosphorylation of Tyr-256 of WASL by LCK N-WASP Phosphorylation Is Enhanced 40-fold upon Binding to Activated Cdc42 wasl-ptpn2-1 2 6 enzyme target enzyme target substrate substrate unspecified role unspecified role gbd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain tyr-256 optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy modified residue ms mass detection of residue modification modified residue ms certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 4 enzyme enzyme unspecified role unspecified role ecoli Escherichia coli (strain K12) dephosphorylation dephosphorylation reaction dephosphorylation Km = 68 uM Kcat = 22 s-1 Fig. 3A, Table 1 Dephosphorylation of Tyr-256 of WASL by PTPN2 Here we examined the effects of the autoinhibited fold on the ability of the T cell phosphatase to dephosphorylate p-Tyr256-N-WASP proteins. Manufacturers website describes this as containing an 11 kDa deletion from the C-terminus of the wild type protein, however this is not described at the residue level and cannot be mapped wasl-ptpn2-2 2 4 enzyme enzyme unspecified role unspecified role ecoli Escherichia coli (strain K12) 3 unspecified role unspecified role ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 enzyme target enzyme target substrate substrate unspecified role unspecified role vca domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain tyr-256 optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy modified residue ms mass detection of residue modification modified residue ms certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig. 3B, Table 1 Presence of CDC42 increased efficiency of dephosphorylation of N-WASP by PTPN2 by 30-fold. Dephosphorylation of Tyr-256 of WASL by PTPN2 N-WASP Dephosphorylation Is Enhanced 30-fold upon Binding to Activated Cdc42 Manufacturers website describes this as containing an 11 kDa deletion from the C-terminus of the wild type protein, however this is not described at the residue level and cannot be mapped