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/ http://www.ebi.ac.uk/ margolis-2010-3 EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Margolis SS., Salogiannis J., Lipton DM., Mandel-Brehm C., Wills ZP., Mardinly AR., Hu L., Greer PL., Bikoff JB., Ho HY., Soskis MJ., Sahin M., Greenberg ME. Apoptosis - Interactions involving proteins with a function related to apoptosis Details of Rhotekin and PAK1 taken from manufacturers website Ephexin B5 mapped using partial sequence given in PMID:15848799 - which gave an exact match to isoform 2 of Q5FWH6. Data on Ube3A could not be captured as this could not be assigned to a single gene product. Michael_Greenberg@hms.harvard.edu Cell (0092-8674) 2010 Only protein-protein interactions imex curation margolis-2010-1 EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. 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 western blot western blot Immuno blot Immuno blot Margolis SS., Salogiannis J., Lipton DM., Mandel-Brehm C., Wills ZP., Mardinly AR., Hu L., Greer PL., Bikoff JB., Ho HY., Soskis MJ., Sahin M., Greenberg ME. Apoptosis - Interactions involving proteins with a function related to apoptosis Ephexin B5 mapped using partial sequence given in PMID:15848799 - which gave an exact match to isoform 2 of Q5FWH6. Data on Ube3A could not be captured as this could not be assigned to a single gene product. Michael_Greenberg@hms.harvard.edu Cell (0092-8674) 2010 Only protein-protein interactions imex curation margolis-2010-2 EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. mouse-brain-cortex Mus musculus brain cortex brain_cortex Brain cortex [Cerebral cortex; Cerebrum cortex; Cortex; Pallium] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Margolis SS., Salogiannis J., Lipton DM., Mandel-Brehm C., Wills ZP., Mardinly AR., Hu L., Greer PL., Bikoff JB., Ho HY., Soskis MJ., Sahin M., Greenberg ME. Apoptosis - Interactions involving proteins with a function related to apoptosis Neurons from embryonic day 16 (E16) mouse brain cortical lysates Ephexin B5 mapped using partial sequence given in PMID:15848799 - which gave an exact match to isoform 2 of Q5FWH6. Data on Ube3A could not be captured as this could not be assigned to a single gene product. Michael_Greenberg@hms.harvard.edu Cell (0092-8674) 2010 Only protein-protein interactions imex curation margolis-2010-4 EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. 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 anti tag western anti tag western blot anti tag western Margolis SS., Salogiannis J., Lipton DM., Mandel-Brehm C., Wills ZP., Mardinly AR., Hu L., Greer PL., Bikoff JB., Ho HY., Soskis MJ., Sahin M., Greenberg ME. Apoptosis - Interactions involving proteins with a function related to apoptosis Ephexin B5 mapped using partial sequence given in PMID:15848799 - which gave an exact match to isoform 2 of Q5FWH6. Data on Ube3A could not be captured as this could not be assigned to a single gene product. Michael_Greenberg@hms.harvard.edu Cell (0092-8674) 2010 Only protein-protein interactions imex curation rtkn_mouse Rhotekin Rtkn protein protein mouse Mus musculus Mouse MFSRNHRSRITVARGSALEMEFKRGRFRLSFFSESPEDTELQRKLDHEIRMRDGACKLLAACSQREQALEATKSLLVCNSRILSYMGELQRRKEAQVLEKTGRRPSDSVQPAQHSPCRGRVCISDLRIPLMWKDTEYFKNKGDLHRWAVFLLLQIGEQIQDTEMVLVDRTLTDISFQNNVLFAEAEPDFELRLELYGACVEEEGALAGAPKRLATKLSSSLGRSSGKRVRASLDSAGASGNSPVLLPTPAVGGPRFHLLAHTTLTLEEVQDGFRTHDLTLTSHEENPAWLPLYGSVCCRLAAQPLCMIQPTASGALRVQQAGELQNGTLVHGVLKGTNLFCYWRSEDADTGQEPLFTIVINKETRVRAGELEQAPEWPFTLSISNKYGDDEVTNTLQLESREALQNWMEALWQLFFDMSQWRHCCDEVMKIETPAPRKPPQALAKQGSLYHEMAIEPLDDIAAVTDILAQREGTRLEPSPPWLAMFTDQPALPSSCSPASVAPVPTWMQPLPWGRPRTFSLDAAPADHSLGPSRSVAPLPPQRSPKSRGFYSKSQLGPWLQSPV rhoa_human Transforming protein RhoA Rho cDNA clone 12 ARHA RHO12 RHOA ARH12 protein protein human Homo sapiens Human MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADIEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKHFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTKDGVREVFEMATRAALQARRGKKKSGCLVL rac1_human Ras-related C3 botulinum toxin substrate 1 RAC1 MIG5 p21-Rac1 Ras-like protein TC25 Cell migration-inducing gene 5 protein TC25 protein protein human Homo sapiens Human MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVMVDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHHCPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQRGLKTVFDEAIRAVLCPPPVKKRKRKCLLL cdc42_human Cell division control protein 42 homolog CDC42 G25K GTP-binding protein protein protein human Homo sapiens Human MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVMIGGEPYTLGLFDTAGQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHHCPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQKGLKNVFDEAILAALEPPEPKKSRRCVLL pak1_human Serine/threonine-protein kinase PAK 1 PAK1 p21-activated kinase 1 p65-PAK Alpha-PAK protein protein human Homo sapiens Human MSNNGLDIQDKPPAPPMRNTSTMIGAGSKDAGTLNHGSKPLPPNPEEKKKKDRFYRSILPGDKTNKKKEKERPEISLPSDFEHTIHVGFDAVTGEFTGMPEQWARLLQTSNITKSEQKKNPQAVLDVLEFYNSKKTSNSQKYMSFTDKSAEDYNSSNALNVKAVSETPAVPPVSEDEDDDDDDATPPPVIAPRPEHTKSVYTRSVIEPLPVTPTRDVATSPISPTENNTTPPDALTRNTEKQKKKPKMSDEEILEKLRSIVSVGDPKKKYTRFEKIGQGASGTVYTAMDVATGQEVAIKQMNLQQQPKKELIINEILVMRENKNPNIVNYLDSYLVGDELWVVMEYLAGGSLTDVVTETCMDEGQIAAVCRECLQALEFLHSNQVIHRDIKSDNILLGMDGSVKLTDFGFCAQITPEQSKRSTMVGTPYWMAPEVVTRKAYGPKVDIWSLGIMAIEMIEGEPPYLNENPLRALYLIATNGTPELQNPEKLSAIFRDFLNRCLEMDVEKRGSAKELLQHQFLKIAKPLSSLTPLIAAAKEATKNNH rhoa_mouse Transforming protein RhoA Rhoa Arha Arha2 protein protein mouse Mus musculus Mouse MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADIEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKHFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTKDGVREVFEMATRAALQARRGKKKSGCLIL ephb2_mouse Ephrin type-B receptor 2 Tyrosine-protein kinase receptor EPH-3 Neural kinase Nuk receptor tyrosine kinase Tyrosine-protein kinase receptor SEK-3 Ephb2 Epth3 Nuk Sek3 protein protein mouse Mus musculus Mouse MAVRRLGAALLLLPLLAAVEETLMDSTTATAELGWMVHPPSGWEEVSGYDENMNTIRTYQVCNVFESSQNNWLRTKFIRRRGAHRIHVEMKFSVRDCSSIPSVPGSCKETFNLYYYEADFDLATKTFPNWMENPWVKVDTIAADESFSQVDLGGRVMKINTEVRSFGPVSRNGFYLAFQDYGGCMSLIAVRVFYRKCPRIIQNGAIFQETLSGAESTSLVAARGSCIANAEEVDVPIKLYCNGDGEWLVPIGRCMCKAGFEAVENGTVCRGCPSGTFKANQGDEACTHCPINSRTTSEGATNCVCRNGYYRADLDPLDMPCTTIPSAPQAVISSVNETSLMLEWTPPRDSGGREDLVYNIICKSCGSGRGACTRCGDNVQYAPRQLGLTEPRIYISDLLAHTQYTFEIQAVNGVTDQSPFSPQFASVNITTNQAAPSAVSIMHQVSRTVDSITLSWSQPDQPNGVILDYELQYYEKELSEYNATAIKSPTNTVTVQGLKAGAIYVFQVRARTVAGYGRYSGKMYFQTMTEAEYQTSIKEKLPLIVGSSAAGLVFLIAVVVIAIVCNRRGFERADSEYTDKLQHYTSGHMTPGMKIYIDPFTYEDPNEAVREFAKEIDISCVKIEQVIGAGEFGEVCSGHLKLPGKREIFVAIKTLKSGYTEKQRRDFLSEASIMGQFDHPNVIHLEGVVTKSTPVMIITEFMENGSLDSFLRQNDGQFTVIQLVGMLRGIAAGMKYLADMNYVHRDLAARNILVNSNLVCKVSDFGLSRFLEDDTSDPTYTSALGGKIPIRWTAPEAIQYRKFTSASDVWSYGIVMWEVMSYGERPYWDMTNQDVINAIEQDYRLPPPMDCPSALHQLMLDCWQKDRNHRPKFGQIVNTLDKMIRNPNSLKAMAPLSSGINLPLLDRTIPDYTSFNTVDEWLEAIKMGQYKESFANAGFTSFDVVSQMMMEDILRVGVTLAGHQKKILNSIQVMRAQMNQIQSVEV q5fwh6-2 Rho guanine nucleotide exchange factor 15 Arhgef15 Kiaa0915 Ephexin-5 protein protein mouse Mus musculus Mouse MSAQSLPAATPPTLKPPRIIRPRPPSRHRAPHSPGPLHNGSSPKALPQISNDASASVCTSIFWEPPTASLKPPALLPPSVSRTSLDSQTSPDSPSSTPSPSPVSRRSISPEPAPCSPVPPPKPSGSSRTPLPSGPTPLQDGSASAPGTVRRLAGKFEWGAEGKAQSSDSLERCSQGSTEVNGEKETPEAALSGNGSQENGTPDAALACPPCCPCVCHVAKPGLELRWVPVGSSEDILRIPCRASPLRASRSRINPPVISHPPVVLTSYRSTAERKLLPPLKPPKPTKVRQDISTSEELPQPDLKLPSEDGIQTATKAWEGDRPEGAPLNAPPVALEGREEEGLDGLKGLQWELPLQDEPLYQTYRAAVLSEELWGVGEDGGPSPANPGEAPTFSRLPGPRNTLWQELPAVRGSGLLESLSPQERRMQESLFEVVTSEASYLRSLRLLTDTFVLSQALRDTLTPRDHHTLFSNVQRVQSVSERFLGTLLSRVRSSPHITDLCDVVHAHAVGPFFVYVDYVRNQQYQEETYSRLMDTNVRFSAELRRLQSLPKCERLPLPSFLLLPFQRITRLRMLLQNILSQTEEGSSRQENAQKALGAVSKIIERCSAEVGRMKQTEELIRLTQRLRFHKVKALPLVSWSRRLELQGELTELGCRRGGVLFTSRPRFTPLCLLLFSDLLLITQPKSGQRLQVLDYAHRSLVQAQQVPDPSGPPTFRLSLLSNHQGRPTHRLLQAASLSDMQRWLGAFPTPGPLPCSPDTIYEDCECSQELCSEPSTPSKTEGQSLESKAPRKHLHKNPEEAEG ngef_mouse Ephexin-1 Eph-interacting exchange protein Neuronal guanine nucleotide exchange factor Ngef protein protein mouse Mus musculus Mouse METKNSEDWGKPQRKSESSSRKSNHGPAEMRPALPPENREAPETGEETQNEEPRRLIPIQRHSLFNRAVRHRHKARSTSERRASDQADLPKMGKSVNERSAFNLPQGRLSPWRTPAQRDTGAQEASESSSTPGNGTTPEECPALTDSPTTLTEALQMIHPIPADSWRNLIEQIGLLYQEYRDKSTLQEIETRRQQDAEIQGNSDGSQVGEDAGEEEEEEEEGEEEELASPPERRALPQICLLSNPHSRFNLWQDLPEIQSSGVLDILQPEEIRLQEAMFELVTSEASYYKSLNLLVSHFMENERLKKILHPSEAHILFSNVLDVMAVSERFLLELEHRMEENIVISDVCDIVYRYAADHFSVYITYVSNQTYQERTYKQLLQEKAAFRELIAQLELDPKCKGLPFSSFLILPFQRITRLKLLVQNILKRVEERSEREGTALDAHKELEMVVKACNEGVRKMSRTEQMISIQKKMEFKIKSVPIISHSRWLLKQGELQQMSGPKTSRTLRTKKLFREIYLFLFNDLLVICRQIPGDKYQVFDSAPRGLLRVEELEDQGQTLANVFILRLLENADDREATYMLKASSQSEMKRWMTSLAPNRRTKFVSFTSRLLDCPQVQCVHPYVAQQPDELTLELADILNILEKTEDGWIFGERLHDQERGWFPSSMTEEILNPKIRSQNLKECFRVHKMEDPQRSQNKDRRKLGSRNRQ epha4_mouse Ephrin type-A receptor 4 Tyrosine-protein kinase receptor SEK-1 Tyrosine-protein kinase receptor MPK-3 Epha4 Sek Sek1 protein protein mouse Mus musculus Mouse MAGIFYFILFSFLFGICDAVTGSRVYPANEVTLLDSRSVQGELGWIASPLEGGWEEVSIMDEKNTPIRTYQVCNVMEASQNNWLRTDWITREGAQRVYIEIKFTLRDCNSLPGVMGTCKETFNLYYYESDNDKERFIRESQFGKIDTIAADESFTQVDIGDRIMKLNTEIRDVGPLSKKGFYLAFQDVGACIALVSVRVFYKKCPLTVRNLAQFPDTITGADTSSLVEVRGSCVNNSEEKDVPKMYCGADGEWLVPIGNCLCNAGHEEQNGECQACKIGYYKALSTDASCAKCPPHSYSVWEGATSCTCDRGFFRADNDAASMPCTRPPSAPLNLISNVNETSVNLEWSSPQNTGGRQDISYNVVCKKCGAGDPSKCRPCGSGVHYTPQQNGLKTTRVSITDLLAHTNYTFEIWAVNGVSKYNPSPDQSVSVTVTTNQAAPSSIALVQAKEVTRYSVALAWLEPDRPNGVILEYEVKYYEKDQNERSYRIVRTAARNTDIKGLNPLTSYVFHVRARTAAGYGDFSEPLEVTTNTVPSRIIGDGANSTVLLVSVSGSVVLVVILIAAFVISRRRSKYSKAKQEADEEKHLNQGVRTYVDPFTYEDPNQAVREFAKEIDASCIKIEKVIGVGEFGEVCSGRLKVPGKREICVAIKTLKAGYTDKQRRDFLSEASIMGQFDHPNIIHLEGVVTKCKPVMIITEYMENGSLDAFLRKNDGRFTVIQLVGMLRGIGSGMKYLSDMSYVHRDLAARNILVNSNLVCKVSDFGMSRVLEDDPEAAYTTRGGKIPIRWTAPEAIAYRKFTSASDVWSYGIVMWEVMSYGERPYWDMSNQDVIKAIEEGYRLPPPMDCPIALHQLMLDCWQKERSDRPKFGQIVNMLDKLIRNPNSLKRTGSESSRPNTALLDPSSPEFSAVVSVGDWLQAIKMDRYKDNFTAAGYTTLEAVVHMSQDDLARIGITAITHQNKILSSVQAMRTQMQQMHGRMVPV rtkn-rhoa-2 Interaction between Rtkn and RhoA 997984 n/a 997988 unspecified role unspecified role bait bait region 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 997989 unspecified role unspecified role prey prey 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. physical association physical association false false false Cells expressing Arhgef15 (E5) exhibited higher levels of activated RhoA compared to cells transfected with a control plasmid, indicating that E5 activates RhoA however cells expressing a form of E5 predicted to be inactive as a GEF, mutated in three conserved amino acids (L562, Q566, and R567) to alanine (E5-LQR) were significantly impaired relative to WT in their ability to activate RhoA. EphB2-mediated tyrosine phosphorylation of E5 triggers a decrease in E5 protein expression resulting in a decrease in RhoA activation. Fig. 2A, 2B, 5F pak1-cdc42-1 Interaction between PAK1 and CDC42 997984 n/a 997990 unspecified role unspecified role prey prey 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. n/a 997991 unspecified role unspecified role prey prey 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. n/a 997992 unspecified role unspecified role bait bait region 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 false pbd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) association association false false false Supp. Fig. 2A Ephexin1-expressing cells displayed enhanced binding of Rac1 and Cdc42 to PAK1 rtkn-rhoa-3 Interaction between Rtkn and RhoA 997984 n/a 997988 unspecified role unspecified role bait bait region 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 997993 unspecified role unspecified role bait bait mouse-brain Mus musculus brain brain Brain [cerebrum] physical association physical association false false false There was a significant decrease in RhoA activation in brain extracts from E5−/− mice compared to wild-type mice, suggesting that E5 is required to maintain wild-type levels of RhoA activity in the brain. Fig. 2C ephb2-arhgef15-1 Interaction between EphB2 and Arhgef15 (E5) 997985 n/a 997994 unspecified role unspecified role bait bait 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 false n/a 997995 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 1B epha4-ngef-1 Interaction between EphA4 and Ngef 997985 n/a 997996 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 997997 unspecified role unspecified role bait bait 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 false physical association physical association false false false Fig. 1B ephb2-arhgef15-2 Interaction between EphB2 and Arhgef15 (E5) 997986 n/a 997994 unspecified role unspecified role prey prey n/a 997995 unspecified role unspecified role bait bait physical association physical association false false false Fig. 1C, Supp. Fig. 1 Interaction not observed in Arhgef15-/- mice ephb2-arhgef15-3 Interaction between EphB2 and Arhgef15 (E5) 997987 n/a 997995 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 997994 unspecified role unspecified role bait bait 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 false cytoplasmic region required to bind necessary binding region required to bind deletion analysis deletion analysis range range certain certain sequence position certain false physical association physical association false false false Supp. Fig. 5A Exact coordinates of cytoplasmic domain not given either in this paper, or that referenced by the article o have been inferred from the UniProtKB entry.