mbinfo MBInfo http://www.mechanobio.info/ Characterization of p190RhoGEF, a RhoA-specific guanine nucleotide exchange factor that interacts with microtubules. Characterization of p190RhoGEF, a RhoA-specific guanine nucleotide exchange factor that interacts with microtubules. J. Biol. Chem. (0021-9258) 2001 imex curation van Horck F.P., Ahmadian M.R., Haeusler L.C., Moolenaar W.H., Kranenburg O. Interaction detection: GST pull-down. The GST pull down assay between p190RhoGEF and GST-RhoA (Fig 3) could not be curated since the source of GST-RhoA was not specified and could not be identified. The colocalization (Fig 6D) and the coimmunoprecipitation (Fig 8) involving tubulins could not be curated since the specific tubulin isoform/subunit involved could not be identified. wmoolen@nki.nl 30-Sept-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot van Horck F.P., Ahmadian M.R., Haeusler L.C., Moolenaar W.H., Kranenburg O. Interaction detection: GST pull-down. The GST pull down assay between p190RhoGEF and GST-RhoA (Fig 3) could not be curated since the source of GST-RhoA was not specified and could not be identified. The colocalization (Fig 6D) and the coimmunoprecipitation (Fig 8) involving tubulins could not be curated since the specific tubulin isoform/subunit involved could not be identified. wmoolen@nki.nl Accepted 2013-SEP-16 by ORCHARD 30-Sept-2013: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation Characterization of p190RhoGEF, a RhoA-specific guanine nucleotide exchange factor that interacts with microtubules. Characterization of p190RhoGEF, a RhoA-specific guanine nucleotide exchange factor that interacts with microtubules. J. Biol. Chem. (0021-9258) 2001 imex curation van Horck F.P., Ahmadian M.R., Haeusler L.C., Moolenaar W.H., Kranenburg O. Interaction detection: GST pull-down. The GST pull down assay between p190RhoGEF and GST-RhoA (Fig 3) could not be curated since the source of GST-RhoA was not specified and could not be identified. The colocalization (Fig 6D) and the coimmunoprecipitation (Fig 8) involving tubulins could not be curated since the specific tubulin isoform/subunit involved could not be identified. wmoolen@nki.nl 30-Sept-2013: Contacted by IntAct-Help. Only protein-protein interactions mouse-n1e-115 Mus musculus neuroblastoma cells n1e-115 Mouse neuroblastoma fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining van Horck F.P., Ahmadian M.R., Haeusler L.C., Moolenaar W.H., Kranenburg O. Interaction detection: GST pull-down. The GST pull down assay between p190RhoGEF and GST-RhoA (Fig 3) could not be curated since the source of GST-RhoA was not specified and could not be identified. The colocalization (Fig 6D) and the coimmunoprecipitation (Fig 8) involving tubulins could not be curated since the specific tubulin isoform/subunit involved could not be identified. wmoolen@nki.nl Accepted 2013-SEP-16 by ORCHARD 30-Sept-2013: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation 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 arg28_mouse Rho guanine nucleotide exchange factor 28 Arhgef28 190 kDa guanine nucleotide exchange factor Kiaa1998 Rgnef Rho guanine nucleotide exchange factor Rho-interacting protein 2 Rhoip2 Rip2 protein protein mouse Mus musculus Mouse MELSCSEVPLYGQKTVYAKFGKNVYLPEDAEFYFVYGGSHQRHVVIADRVQDNVLQSSIPGHWLQETVTVSVCLCSEGYSPVTMGSGSVTYVDNMACRLARLLVTQADRLTACSHQTLLTPFALTVEALPALDEELVLALTQLELPLGWTVLGNSSLEVSLHRESLLHLAVRWALPKLFHFLLCLPGGVKALKLPNEEATTPLDLALQGGHSTLVEDITNFQGSHSPGFSRLRLNEEATLQFVHSSETLTLTVNHTAEHLLEADIKLFRKYFWDRAFLVKALEQEAKTEKATMPSGAAETEEEVRNLESGRSPSEEEEDAKSIKSQVDGPSEHEDQDRLALDRSFDGLKKSKHVPASLAAGQLSDVLNGGDEVYANCMVIDQVGDLDINYINLEGLSTHTSPESGRSMLGPQACMHTLPPDTSPCGRPLIENSEGTLDAAASQSFVTPSSSRTSNLNLSFGLHGFEKEQSHLKKRSSSLDALVADSEGEGGSEPPICYAVGSQSSPRTGLPSGDELDSFETNTEPDCNISRTESLSLSSTLHSKESLLSGIRSRSYSCSSPKISSGKSRLVRDFTVCSTSEEQRSYSFQEPPGEKRIQEEEWDEYVIPAKSESEKYKVSRTFSFLMNRMTSPRNKSKMKNKDTKEKEKMNRHQFVPGTFSGVLQCSGCDKTLLGKESLQCANCKANTHKGCKDAVPPCTKKFQEKYNKNKPQSILGSSSVRDVPAPGLSLHPSSSMPIGLPAGRKEFAAQVHPLSRSVPGTTLESFRRAVTSLESEGDSWRSRSHSDELFQSMGSSPSTESFMMEDVVDSSLWIDLSSDAQEFEAESWSLVVDPSFCSRQEKDVIKRQDVIFELMQTEVHHIQTLLIMSEVFRKGMKEELQLDHSTVDKIFPCLDELLETHRHFFFSMKERRQESCAGSDRNFVINQIGDILVQQFSEENASKMKRIYGEFCSHHKEAMSLFKELQQNKKFQNFIKIRNSNLLARRRGIPECILLVTQRITKYPVLVERILQYTKERTEEHRDLCKALGLIKDMIAAVDLKVSEYEKNQKWLEILNKIENKTYTKLKNGHVFRKQALLSQERALLHDGLVYWKTATGRFKDILALLLTDVLLFLQEKDQKYIFAAVDQKPSVISLQKLIAREVANEERGMFLISASSAGPEMYEIHTNSKEERNNWMRRIQQAVESCPEEEGGRTSESDEERRKAEARVAKIQQCQEILSNQDQQICTYLEEKLHIYAELGELSGFEDVHLEPHLLIKPDPGEPPQAASLLAAALREAESLQVAVKASKMGDVSQSSEESPGGTVLMDTPSTQDVPASPTASLVTEGTEGRGCWDVDPGLQGVVTDLAVSDAGEKVEYRSFSGSSQSEIIQAIQNLTRLLYSLQAALTIQDSHIEIHKLVLQQRESLAPSHSFRGGPLQDQEKSRYLEKQREELANIHKLQHQFQQEQRRWHRTCDQQQREQEAQESWLQARERECQSQEELLLRHRSELDHQLQEYQQSLERLREGQRMVERERQKMRVQQGLLGHCKHSRQRSLPAVFSPGSKEVTELNRAESLCHENSFFINEAFGHMSLNTSNKPNPSGVPWDAHPLEGSHFDLARTSESPTELKIDISQPPDVNSELWTTGPGHQRPALQENSKESYKNVADLDSFQSESSSPQDSNQRWPPATDTHNRSKTKSSDGGWTRWRCWRRG mint Guanine nucleotide exchange factor that interacts with RHOA, but not with RAC or CDC42. Activates RHOA to promote cytoskeletal contraction and inhibit neurite outgrowth 4E96087C449FF14C rhoa_chlae_protein RhoA protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey 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_chlae_protein Rac1 protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey 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 rtkn-rhoa-11 1 5 unspecified role unspecified role prey prey chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 8 unspecified role unspecified role bait bait c21 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain The amino acid range for the C21 domain of Rhotekin [7-113], was inferred from the reference from which the organism source of Rhotekin was traced. 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 ecoli Escherichia coli (strain K12) physical association physical association Fig 2B, 9. GST-Rhotekin precipitated only activated (GTP-loaded) RhoA. RhoA was activated only in cells transfected with p190RhoGEF-DH/PH and not in cells transfected with Tiam1 (Fig 2B). RhoA was activated in cells treated with full length p190RhoGEF and in cells treated with nocodazole (causes microtubule disruption), but there was no enhanced RhoGEF-mediated RhoA activation in microtubule-disrupted cells (Fig 9). pak1-rac1-19 1 7 unspecified role unspecified role prey prey chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 3 unspecified role unspecified role bait bait crib domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain The amino acid range for the Pak1b CRIB domain [56-272] was inferred from the reference from which the organism source of PAK was traced. 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 ecoli Escherichia coli (strain K12) physical association physical association Fig 2B. GST-PAK-CRIB precipitated only activated (GTP-loaded) Rac1. Rac1 was activated only in cells transfected with Tiam1 and not in cells transfected with p190RhoGEF-DH/PH. arhgef28-rhoa 2 4 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig 6B.