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/ EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. Cell (0092-8674) 2010 imex curation 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 06-April-2011: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Accepted 2010-Not a month!-8 by JYOTI. 06-April-2011: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions imex curation EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. Cell (0092-8674) 2010 imex curation 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 06-April-2011: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Accepted 2010-Not a month!-8 by JYOTI. 06-April-2011: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions imex curation EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. Cell (0092-8674) 2010 imex curation 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 06-April-2011: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Accepted 2010-Not a month!-8 by JYOTI. 06-April-2011: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions imex curation EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. EphB-mediated degradation of the RhoA GEF Ephexin5 relieves a developmental brake on excitatory synapse formation. Cell (0092-8674) 2010 imex curation 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 06-April-2011: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Accepted 2010-Not a month!-8 by JYOTI. 06-April-2011: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti 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 pak1_human Serine/threonine-protein kinase PAK 1 PAK1 p21-activated kinase 1 p65-PAK Alpha-PAK protein protein human Homo sapiens Human MSNNGLDIQDKPPAPPMRNTSTMIGAGSKDAGTLNHGSKPLPPNPEEKKKKDRFYRSILPGDKTNKKKEKERPEISLPSDFEHTIHVGFDAVTGEFTGMPEQWARLLQTSNITKSEQKKNPQAVLDVLEFYNSKKTSNSQKYMSFTDKSAEDYNSSNALNVKAVSETPAVPPVSEDEDDDDDDATPPPVIAPRPEHTKSVYTRSVIEPLPVTPTRDVATSPISPTENNTTPPDALTRNTEKQKKKPKMSDEEILEKLRSIVSVGDPKKKYTRFEKIGQGASGTVYTAMDVATGQEVAIKQMNLQQQPKKELIINEILVMRENKNPNIVNYLDSYLVGDELWVVMEYLAGGSLTDVVTETCMDEGQIAAVCRECLQALEFLHSNQVIHRDIKSDNILLGMDGSVKLTDFGFCAQITPEQSKRSTMVGTPYWMAPEVVTRKAYGPKVDIWSLGIMAIEMIEGEPPYLNENPLRALYLIATNGTPELQNPEKLSAIFRDFLNRCLEMDVEKRGSAKELLQHQFLKIAKPLSSLTPLIAAAKEATKNNH DIP protein Q13153 original sequence version: 161 DIP protein Q13153 original sequence version: 122 DIP protein Q13153 original sequence version: 134 DIP protein Q13153 original sequence version: 165 1A95CD5F2195CD7B rhoa_human Transforming protein RhoA Rho cDNA clone 12 ARHA RHO12 RHOA ARH12 protein protein human Homo sapiens Human MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADIEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKHFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTKDGVREVFEMATRAALQARRGKKKSGCLVL DIP protein P61586 original sequence version: 86 DIP protein P61586 original sequence version: 123 DIP protein P61586 original sequence version: 57 DIP protein P61586 original sequence version: 88 DIP protein P61586 original sequence version: 87 DIP protein P61586 original sequence version: 85 DIP protein P61586 original sequence version: 125 DIP protein P61586 original sequence version: 132 C4DA2DC31FF858BC q5fwh6-2 Rho guanine nucleotide exchange factor 15 Arhgef15 Kiaa0915 Ephexin-5 protein protein mouse Mus musculus Mouse MSAQSLPAATPPTLKPPRIIRPRPPSRHRAPHSPGPLHNGSSPKALPQISNDASASVCTSIFWEPPTASLKPPALLPPSVSRTSLDSQTSPDSPSSTPSPSPVSRRSISPEPAPCSPVPPPKPSGSSRTPLPSGPTPLQDGSASAPGTVRRLAGKFEWGAEGKAQSSDSLERCSQGSTEVNGEKETPEAALSGNGSQENGTPDAALACPPCCPCVCHVAKPGLELRWVPVGSSEDILRIPCRASPLRASRSRINPPVISHPPVVLTSYRSTAERKLLPPLKPPKPTKVRQDISTSEELPQPDLKLPSEDGIQTATKAWEGDRPEGAPLNAPPVALEGREEEGLDGLKGLQWELPLQDEPLYQTYRAAVLSEELWGVGEDGGPSPANPGEAPTFSRLPGPRNTLWQELPAVRGSGLLESLSPQERRMQESLFEVVTSEASYLRSLRLLTDTFVLSQALRDTLTPRDHHTLFSNVQRVQSVSERFLGTLLSRVRSSPHITDLCDVVHAHAVGPFFVYVDYVRNQQYQEETYSRLMDTNVRFSAELRRLQSLPKCERLPLPSFLLLPFQRITRLRMLLQNILSQTEEGSSRQENAQKALGAVSKIIERCSAEVGRMKQTEELIRLTQRLRFHKVKALPLVSWSRRLELQGELTELGCRRGGVLFTSRPRFTPLCLLLFSDLLLITQPKSGQRLQVLDYAHRSLVQAQQVPDPSGPPTFRLSLLSNHQGRPTHRLLQAASLSDMQRWLGAFPTPGPLPCSPDTIYEDCECSQELCSEPSTPSKTEGQSLESKAPRKHLHKNPEEAEG A1CA2F81739D3073 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 DIP protein P54763 original sequence version: 151 DIP protein P54763 original sequence version: 119 DIP protein P54763 original sequence version: 123 92DB5234F18758A7 rtkn_mouse Rhotekin Rtkn protein protein mouse Mus musculus Mouse MFSRNHRSRITVARGSALEMEFKRGRFRLSFFSESPEDTELQRKLDHEIRMRDGACKLLAACSQREQALEATKSLLVCNSRILSYMGELQRRKEAQVLEKTGRRPSDSVQPAQHSPCRGRVCISDLRIPLMWKDTEYFKNKGDLHRWAVFLLLQIGEQIQDTEMVLVDRTLTDISFQNNVLFAEAEPDFELRLELYGACVEEEGALAGAPKRLATKLSSSLGRSSGKRVRASLDSAGASGNSPVLLPTPAVGGPRFHLLAHTTLTLEEVQDGFRTHDLTLTSHEENPAWLPLYGSVCCRLAAQPLCMIQPTASGALRVQQAGELQNGTLVHGVLKGTNLFCYWRSEDADTGQEPLFTIVINKETRVRAGELEQAPEWPFTLSISNKYGDDEVTNTLQLESREALQNWMEALWQLFFDMSQWRHCCDEVMKIETPAPRKPPQALAKQGSLYHEMAIEPLDDIAAVTDILAQREGTRLEPSPPWLAMFTDQPALPSSCSPASVAPVPTWMQPLPWGRPRTFSLDAAPADHSLGPSRSVAPLPPQRSPKSRGFYSKSQLGPWLQSPV DIP protein Q8C6B2 original sequence version: 41 DIP protein Q8C6B2 original sequence version: 57 0934EEBF09B12A3E rhoa_mouse Transforming protein RhoA Rhoa Arha Arha2 protein protein mouse Mus musculus Mouse MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADIEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKHFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTKDGVREVFEMATRAALQARRGKKKSGCLIL DIP protein Q9QUI0 original sequence version: 76 DIP protein Q9QUI0 original sequence version: 77 DIP protein Q9QUI0 original sequence version: 100 C4C8BDC31FF858BC 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 ACEDF83A45E5EA67 ngef_mouse Ephexin-1 Eph-interacting exchange protein Neuronal guanine nucleotide exchange factor Ngef protein protein mouse Mus musculus Mouse METKNSEDWGKPQRKSESSSRKSNHGPAEMRPALPPENREAPETGEETQNEEPRRLIPIQRHSLFNRAVRHRHKARSTSERRASDQADLPKMGKSVNERSAFNLPQGRLSPWRTPAQRDTGAQEASESSSTPGNGTTPEECPALTDSPTTLTEALQMIHPIPADSWRNLIEQIGLLYQEYRDKSTLQEIETRRQQDAEIQGNSDGSQVGEDAGEEEEEEEEGEEEELASPPERRALPQICLLSNPHSRFNLWQDLPEIQSSGVLDILQPEEIRLQEAMFELVTSEASYYKSLNLLVSHFMENERLKKILHPSEAHILFSNVLDVMAVSERFLLELEHRMEENIVISDVCDIVYRYAADHFSVYITYVSNQTYQERTYKQLLQEKAAFRELIAQLELDPKCKGLPFSSFLILPFQRITRLKLLVQNILKRVEERSEREGTALDAHKELEMVVKACNEGVRKMSRTEQMISIQKKMEFKIKSVPIISHSRWLLKQGELQQMSGPKTSRTLRTKKLFREIYLFLFNDLLVICRQIPGDKYQVFDSAPRGLLRVEELEDQGQTLANVFILRLLENADDREATYMLKASSQSEMKRWMTSLAPNRRTKFVSFTSRLLDCPQVQCVHPYVAQQPDELTLELADILNILEKTEDGWIFGERLHDQERGWFPSSMTEEILNPKIRSQNLKECFRVHKMEDPQRSQNKDRRKLGSRNRQ F255DE351E02A586 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 DIP protein Q03137 original sequence version: 161 DIP protein Q03137 original sequence version: 101 89BEB2C7CDB54A55 ephb2-arhgef15-1 3 9 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 8 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 physical association physical association We found that E5 coimmunoprecipitates with EphB2 Fig. 1B epha4-ngef-1 3 13 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 14 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 physical association physical association Fig. 1B By contrast, E1 is coimmunoprecipitated by EphA4 ephb2-arhgef15-2 4 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association This analysis revealed that endogenous, neuronal EphB2 is immunoprecipitated by α-C-E5 but not IgG. Fig. 1C, Supp. Fig. 1 Interaction not observed in Arhgef15-/- mice rtkn-rhoa-2 1 10 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 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 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 We prepared extracts from the transfected cells and incubated the extracts with a GST-fusion protein that includes the Rhotekin-Binding Domain (GST-RBD), a protein domain that selectively interacts with active (GTP-bound) but not inactive (GDP-bound) RhoA. pak1-cdc42-1 1 12 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. 5 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. 6 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 pbd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) association association Supp. Fig. 2A Ephexin1-expressing cells displayed enhanced binding of Rac1 and Cdc42 to PAK1 rtkn-rhoa-3 1 10 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role bait bait mouse-brain Mus musculus brain brain Brain [cerebrum] physical association physical association 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-arhgef15-3 2 8 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 9 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 cytoplasmic region required to bind necessary binding region required to bind deletion analysis deletion analysis range range certain certain sequence position certain physical association physical association We found that E5 coimmunoprecipitates with EphB2 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.