mbinfo MBInfo http://www.mechanobio.info/ http://www.mechanobio.info/ oser-2009-2 Cortactin regulates cofilin and N-WASp activities to control the stages of invadopodium assembly and maturation. rat-mtln3 Rat mammary adenocarcinoma mtln3 Rat mammary adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Oser M., Yamaguchi H., Mader C.C., Bravo-Cordero J.J., Arias M., Chen X., Desmarais V., van Rheenen J., Koleske A.J., Condeelis J. Interaction detection: FRET acceptor photobleaching. The FRET experiments from Fig S4(C,D) could not be curated since the source of GFP-cofilin could not be identified. The cortactin-Nck1 colocalization experiments (Fig 7C) could not be curated since the source of GFP-Nck1 could not be identified. moser@aecom.yu.edu J. Cell Biol. (0021-9525) 2009 Only protein-protein interactions imex curation oser-2009-1 Cortactin regulates cofilin and N-WASp activities to control the stages of invadopodium assembly and maturation. rat-mtln3 Rat mammary adenocarcinoma mtln3 Rat mammary adenocarcinoma fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret immunostaining immunostaining Oser M., Yamaguchi H., Mader C.C., Bravo-Cordero J.J., Arias M., Chen X., Desmarais V., van Rheenen J., Koleske A.J., Condeelis J. Interaction detection: FRET acceptor photobleaching. The FRET experiments from Fig S4(C,D) could not be curated since the source of GFP-cofilin could not be identified. The cortactin-Nck1 colocalization experiments (Fig 7C) could not be curated since the source of GFP-Nck1 could not be identified. moser@aecom.yu.edu J. Cell Biol. (0021-9525) 2009 Only protein-protein interactions imex curation oser-2009-3 Cortactin regulates cofilin and N-WASp activities to control the stages of invadopodium assembly and maturation. in vitro In vitro affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom weight by comassie molecular weight estimation by coomasie staining weight by comassie Oser M., Yamaguchi H., Mader C.C., Bravo-Cordero J.J., Arias M., Chen X., Desmarais V., van Rheenen J., Koleske A.J., Condeelis J. Interaction detection: FRET acceptor photobleaching. The FRET experiments from Fig S4(C,D) could not be curated since the source of GFP-cofilin could not be identified. The cortactin-Nck1 colocalization experiments (Fig 7C) could not be curated since the source of GFP-Nck1 could not be identified. moser@aecom.yu.edu J. Cell Biol. (0021-9525) 2009 Only protein-protein interactions imex curation oser-2009-4 Cortactin regulates cofilin and N-WASp activities to control the stages of invadopodium assembly and maturation. rat-mtln3 Rat mammary adenocarcinoma mtln3 Rat mammary adenocarcinoma anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Oser M., Yamaguchi H., Mader C.C., Bravo-Cordero J.J., Arias M., Chen X., Desmarais V., van Rheenen J., Koleske A.J., Condeelis J. Interaction detection: FRET acceptor photobleaching. The FRET experiments from Fig S4(C,D) could not be curated since the source of GFP-cofilin could not be identified. The cortactin-Nck1 colocalization experiments (Fig 7C) could not be curated since the source of GFP-Nck1 could not be identified. moser@aecom.yu.edu J. Cell Biol. (0021-9525) 2009 Only protein-protein interactions imex curation oser-2009-5 Cortactin regulates cofilin and N-WASp activities to control the stages of invadopodium assembly and maturation. rat-mtln3 Rat mammary adenocarcinoma mtln3 Rat mammary adenocarcinoma fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Oser M., Yamaguchi H., Mader C.C., Bravo-Cordero J.J., Arias M., Chen X., Desmarais V., van Rheenen J., Koleske A.J., Condeelis J. Interaction detection: FRET acceptor photobleaching. The FRET experiments from Fig S4(C,D) could not be curated since the source of GFP-cofilin could not be identified. The cortactin-Nck1 colocalization experiments (Fig 7C) could not be curated since the source of GFP-Nck1 could not be identified. moser@aecom.yu.edu J. Cell Biol. (0021-9525) 2009 Only protein-protein interactions imex curation src8_rat Src substrate cortactin Cttn protein protein rat Rattus norvegicus (Rat) Rat MWKASAGHAVSITQDDGGADDWETDPDFVNDVSEKEQRWGAKTVQGSGHQEHINIHKLRENVFQEHQTLKEKELETGPKASHGYGGKFGVEQDRMDKSAVGHEYQSKLSKHCSQVDSVRGFGGKFGVQMDRVDQSAVGFEYQGKTEKHASQKDYSSGFGGKYGVQADRVDKSAVGFDYQGKTEKHESQKDYSKGFGGKYGIDKDKVDKSAVGFEYQGKTEKHESQKDYVKGFGGKFGVQTDRQDKCALGWDHQEKLQLHESQKDYAKGFGGKYGVQKDRMDKNASTFEEVVQVPSAYQKTVPIEAVTSKTSNIRANFENLAKEREQEDRRKAEAERAQRMAQERQEQEEARRKLEEQARAKKQTPPASPSPQPAEDRPPSSPIYEDAAPLKAEPSYGSSEPEPEYSTEAAGLPEASNQQGLAYTSEPVYETTEVPGHYQAEDDTYDGYESDLGITAIALYDYQAAGDDEISFDPDDVITNIEMIDDGWWRGVCKGRYGLFPANYVELRQ cof1_rat Cofilin-1 Cofilin, non-muscle isoform Cfl1 protein protein rat Rattus norvegicus (Rat) Rat MASGVAVSDGVIKVFNDMKVRKSSTPEEVKKRKKAVLFCLSEDKKNIILEEGKEILVGDVGQTVDDPYTTFVKMLPDKDCRYALYDATYETKESKKEDLVFIFWAPESAPLKSKMIYASSKDAIKKKLTGIKHELQANCYEEVKDRCTLAEKLGGSAVISLEGKPL src8_mouse Src substrate cortactin Cttn Ems1 protein protein mouse Mus musculus Mouse MWKASAGHAVSITQDDGGADDWETDPDFVNDVSEKEQRWGAKTVQGSGHQEHINIHKLRENVFQEHQTLKEKELETGPKASHGYGGKFGVEQDRMDRSAVGHEYQSKLSKHCSQVDSVRGFGGKFGVQMDRVDQSAVGFEYQGKTEKHASQKDYSSGFGGKYGVQADRVDKSAVGFDYQGKTEKHESQKDYSKGFGGKYGIDKDKVDKSAVGFEYQGKTEKHESQKDYVKGFGGKFGVQTDRQDKCALGWDHQEKLQLHESQKDYKTGFGGKFGVQSERQDSSAVGFDYKERLAKHESQQDYAKGFGGKYGVQKDRMDKNASTFEEVVQVPSAYQKTVPIEAVTSKTSNIRANFENLAKEREQEDRRKAEAERAQRMAKERQEQEEARRKLEEQARAKKQTPPASPSPQPIEDRPPSSPIYEDAAPFKAEPSYRGSEPEPEYSIEAAGIPEAGSQQGLTYTSEPVYETTEAPGHYQAEDDTYDGYESDLGITAIALYDYQAAGDDEISFDPDDIITNIEMIDDGWWRGVCKGRYGLFPANYVELRQ b2rz33_rat Cytoplasmic protein Nck1 Nck1_predicted rCG_25905 protein protein rat Rattus norvegicus (Rat) Rat MAEEVVVVAKFDYVAQQEQELDIKKNERLWLLDDSKSWWRVRNSMNKTGFVPSNYVERKNSARKASIVKNLKDTLGIGKVKRKPSVPDTASPADDSFVDPGERLYDLNMPAFVKFNYMAEREDELSLIKGTKVIVMEKCSDGWWRGSYNGQIGWFPSNYVTEEGDSPLGDHVGSLSEKLAAVVNNLNTGQVLHVVQALYPFSSSNDEELNFEKGDVMDVIEKPENDPEWWKCRKINGMVGLVPKNYVTVMQNNPLTSGLEPSPPQCDYIRPSLTGKFAGNPWYYGKVTRHQAEMALNERGHEGDFLIRDSESSPNDFSVSLKAQGKNKHFKVQLKETVYCIGQRKFSTMEELVEHYKKAPIFTSEQGEKLYLVKHLS actb_rat Actin, cytoplasmic 1 Actb Beta-actin protein protein rat Rattus norvegicus (Rat) Rat MDDDIAALVVDNGSGMCKAGFAGDDAPRAVFPSIVGRPRHQGVMVGMGQKDSYVGDEAQSKRGILTLKYPIEHGIVTNWDDMEKIWHHTFYNELRVAPEEHPVLLTEAPLNPKANREKMTQIMFETFNTPAMYVAIQAVLSLYASGRTTGIVMDSGDGVTHTVPIYEGYALPHAILRLDLAGRDLTDYLMKILTERGYSFTTTAEREIVRDIKEKLCYVALDFEQEMATAASSSSLEKSYELPDGQVITIGNERFRCPEALFQPSFLGMESCGIHETTFNSIMKCDVDIRKDLYANTVLSGGTTMYPGIADRMQKEITALAPSTMKIKIIAPPERKYSVWIGGSILASLSTFQQMWISKQEYDESGPSIVHRKCF cofa_dicdi Cofilin-1A cofA cof1 dcof1 DDB_G0277833 protein protein dicdi Dictyostelium discoideum (Slime mold) Slime mold MSSGIALAPNCVSTFNDLKLGRKYGGIIYRISDDSKEIIVDSTLPAGCSFDEFTKCLPENECRYVVLDYQYKEEGAQKSKICFVAWCPDTANIKKKMMATSSKDSLRKACVGIQVEIQGTDASEVKDSCFYEKCTKI wasl_rat Neural Wiskott-Aldrich syndrome protein Wasl protein protein rat Rattus norvegicus (Rat) Rat MSSGQQPPRRVTNVGSLLLTPQENESLFSFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQVALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDAPNGPNLPMATVDIKNPEITTNRFYSSQVNNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELKNLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPPHSSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPGVVVPPPPPNRMYPPPPPALPSSAPSGPPPPPPLSMAGSTAPPPPPPPPPPPGPPPPPGLPSDGDHQVPASSGNKAALLDQIREGAQLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVSDGQESTPPTPAPTSGIVGALMEVMQKRSKAIHSSDEDEDDDDEEDFQDDDEWED cfl1-cttn-5 1372734 n/a 1372739 unspecified role unspecified role prey prey n/a 1372740 unspecified role unspecified role bait bait physical association physical association false false false Fig 5D, 5E. Cortactin coprecipitation by cofilin was decreased in the presence of EGF stimulation. Cofilin did not interact with phosphorylated cortactin. cttn-cfl1 1372734 n/a 1372740 unspecified role unspecified role prey prey n/a 1372739 unspecified role unspecified role bait bait physical association physical association false false false Fig. 5F, 5G The amount of cofilin coprecipitated by cortactin decreased in the presence of EGF stimulation. cfl1-cttn-7 1372734 n/a 1372741 unspecified role unspecified role prey prey Q60598:p.[Tyr421Phe;Tyr466Phe;Tyr482Phe] mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n/a 1372740 unspecified role unspecified role bait bait physical association physical association false false false Fig 5H, 5I. Cortactin 3YF mutant coprecipitated increased levels of cofilin, when compared to WT cortactin, suggesting that cofilin interacts chiefly with dephosphorylated cortactin. nck1-cttn 1372734 n/a 1372739 unspecified role unspecified role prey prey tyr421 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 certain certain sequence position certain certain certain sequence position certain false Required phosphorylation n/a 1372742 unspecified role unspecified role bait bait physical association physical association false false false Fig 7E. Nck1 coprecipitated phosphorylated cortactin as detected by anti-phospho antibody. Nck1 did not coprecipitate non-phosphorylated cortactin. cfl1-cttn-1 1372735 n/a 1372740 unspecified role unspecified role neutral component neutral component n/a 1372739 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false Fig 5A, 5B. The FRET efficiency was increased in starved MLTn3 cells, decreased 1 min after EGF stimulation and increased again after 5 min of EGF stimulation (Fig 5B). cfl1-cttn-3 1372735 n/a 1372740 unspecified role unspecified role neutral component neutral component n/a 1372741 unspecified role unspecified role neutral component neutral component Q60598:p.[Tyr421Glu;Tyr466Glu;Tyr482Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false proximity proximity close-contact colocalization neighbourhood interaction false false false Fig 5C. actb-cfl1-1 1372735 n/a 1372740 unspecified role unspecified role neutral component neutral component n/a 1372743 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false Fig 6F, 6G. The FRET efficiency between cofilin and beta-actin increased in the presence of EGF stimulation. cttn-cofa 1372736 n/a 1372741 unspecified role unspecified role bait bait Q60598:p.[Tyr421Phe;Tyr466Phe;Tyr482Phe] mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 n/a 1372744 unspecified role unspecified role prey prey dicdi Dictyostelium discoideum (Slime mold) Slime mold direct interaction direct interaction false false false 1372736 Fig 6A, 6B. The recombinant cofilin had a higher affinity for the 3YF cortactin mutant, when compared to the WT cortactin, as inferred from the Kd values. The Kd value for the cofilin-cortactin WT interaction was 3.5+/_ 1.1 micromoles and the Kd value for the cofilin-cortactin 3YF mutant was 2.8+/_ 0.34 micromoles. cttn-cfl1-2 1372737 n/a 1372739 unspecified role unspecified role bait bait Q66HL2:p.[Leu421Phe;Gly466Phe;Glu482Phe] mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1372740 unspecified role unspecified role prey prey physical association physical association false false false Fig S5(B,C). wasl-1 1372738 n/a 1372745 unspecified role unspecified role self self n-terminal region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false c-terminal region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false self interaction self interaction false true false Fig 7F, 7G. The intramolecular interaction within N-Wasp was decreased in MTLn3 cells expressing 3YF cortactin, but not in cells expressing WT cortactin.