mbinfo MBInfo http://www.mechanobio.info/ http://www.mechanobio.info/ mcniven-2000-1 Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot McNiven M.A., Kim L., Krueger E.W., Orth J.D., Cao H., Wong T.W. The source of cortactin and dynamin-2 (both endogenous) inferred from the cells (NIH/3T3) used. Contact-email not given in the paper. J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation mcniven-2000-2 Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot McNiven M.A., Kim L., Krueger E.W., Orth J.D., Cao H., Wong T.W. Interaction detection: GST pull down. The source of cortactin and dynamin-2 (both endogenous) inferred from the cells (NIH/3T3) used. Contact-email not given in the paper. J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation mcniven-2000-3 Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting anti tag western anti tag western blot anti tag western McNiven M.A., Kim L., Krueger E.W., Orth J.D., Cao H., Wong T.W. The source of cortactin and dynamin-2 (both endogenous) inferred from the cells (NIH/3T3) used. Contact-email not given in the paper. J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation mcniven-2000-4 Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining McNiven M.A., Kim L., Krueger E.W., Orth J.D., Cao H., Wong T.W. Endogenous proteins in untransfected cells were detected by indirect immunocytochemistry by immunostaining them with primary and fluorescently-labeled secondary antibodies. Cells trasfected with fluorescently-labeled proteins were visualized by tag fluorescence. The source of cortactin and dynamin-2 (both endogenous) inferred from the cells (NIH/3T3) used. Contact-email not given in the paper. J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation dyn2_mouse Dynamin-2 Dynamin UDNM Dnm2 Dyn2 protein protein mouse Mus musculus Mouse MGNRGMEELIPLVNKLQDAFSSIGQSCHLDLPQIAVVGGQSAGKSSVLENFVGRDFLPRGSGIVTRRPLILQLIFSKTEYAEFLHCKSKKFTDFDEVRQEIEAETDRVTGTNKGISPVPINLRVYSPHVLNLTLIDLPGITKVPVGDQPPDIEYQIKDMILQFISRESSLILAVTPANMDLANSDALKLAKEVDPQGLRTIGVITKLDLMDEGTDARDVLENKLLPLRRGYIGVVNRSQKDIEGKKDIRAALAAERKFFLSHPAYRHMADRMGTPHLQKTLNQQLTNHIRESLPTLRSKLQSQLLSLEKEVEEYKNFRPDDPTRKTKALLQMVQQFGVDFEKRIEGSGDQVDTLELSGGARINRIFHERFPFELVKMEFDEKDLRREISYAIKNIHGVRTGLFTPDLAFEAIVKKQVVKLKEPCLKCVDLVIQELISTVRQCTSKLSSYPRLREETERIVTTYIREREGRTKDQILLLIDIEQSYINTNHEDFIGFANAQQRSTQLNKKRAIPNQGEILVIRRGWLTINNISLMKGGSKEYWFVLTAESLSWYKDEEEKEKKYMLPLDNLKIRDVEKGFMSNKHVFAIFNTEQRNVYKDLRQIELACDSQEDVDSWKASFLRAGVYPEKDQAENEDGAQENTFSMDPQLERQVETIRNLVDSYVAIINKSIRDLMPKTIMHLMINNTKAFIHHELLAYLYSSADQSSLMEESAEQAQRRDDMLRMYHALKEALNIIGDISTSTVSTPVPPPVDDTWLQNTSGHSPTPQRRPVSSVHPPGRPPAVRGPTPGPPLIPMPVGATSSFSAPPIPSRPGPQSVFANNDPFSAPPQIPSRPARIPPGIPPGVPSRRAPAAPSRPTIIRPAEPSLLD src8_mouse Src substrate cortactin Cttn Ems1 protein protein mouse Mus musculus Mouse MWKASAGHAVSITQDDGGADDWETDPDFVNDVSEKEQRWGAKTVQGSGHQEHINIHKLRENVFQEHQTLKEKELETGPKASHGYGGKFGVEQDRMDRSAVGHEYQSKLSKHCSQVDSVRGFGGKFGVQMDRVDQSAVGFEYQGKTEKHASQKDYSSGFGGKYGVQADRVDKSAVGFDYQGKTEKHESQKDYSKGFGGKYGIDKDKVDKSAVGFEYQGKTEKHESQKDYVKGFGGKFGVQTDRQDKCALGWDHQEKLQLHESQKDYKTGFGGKFGVQSERQDSSAVGFDYKERLAKHESQQDYAKGFGGKYGVQKDRMDKNASTFEEVVQVPSAYQKTVPIEAVTSKTSNIRANFENLAKEREQEDRRKAEAERAQRMAKERQEQEEARRKLEEQARAKKQTPPASPSPQPIEDRPPSSPIYEDAAPFKAEPSYRGSEPEPEYSIEAAGIPEAGSQQGLTYTSEPVYETTEAPGHYQAEDDTYDGYESDLGITAIALYDYQAAGDDEISFDPDDIITNIEMIDDGWWRGVCKGRYGLFPANYVELRQ src8_chick Src substrate protein p85 CTTN1 EMS1 P85.25 p80 Cortactin protein protein chick Gallus gallus (Chicken) Chicken MTVLLLVVLQMWKATAGHSIAVSQDDGADDWETDPDFVNDVSEKEQRWGAKTVKGSGHQEHINIHQLRENVFQEHQTIKEKELETGPKASHGYGGKFGVEQDRMDKSAVGHEYQSKLSKHCSQVDSVKGFGGKFGVQTDRVDQSAVGFEYQGKTEKHASQKDYSSGFGGKYGVQADRVDKSAVGFDYQGKTEKHESQKDYSKGFGGKYGVDKDKVDKSAVGFEYQGKTEKHESQKDYVKGFGGKFGVQTDRQDKCALGWDHQEKVQLHESQKDYKSGFGGKFGVQTERQDPSAVGFDYKEKLAKHESQQDYSKGFGGKYGVQKDRMDKNAATFEDIEKPTSTYQKTKPVERVANKTSSIRANLENLAKEKEQEDRRKAEAERAQRMAREKQEQEEARRKLEEQAKAKKQTPPPSPTTQPAEPKTPSSPVYQDAVSYDAESAYKNSSTTYSAEHEPESGYKTTGSDYQEAVSQREAEYEPETVYEVAGAGDHYQAEENTYDEYENELGITAIALYDYQAAGDDEISFDPDDIITNIEMIDDGWWRGVCKGRYGLFPANYVELRQ plcg1_human 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-1 PLC-148 PLC1 Phospholipase C-II PLCG1 Phosphoinositide phospholipase C-gamma-1 Phospholipase C-gamma-1 protein protein human Homo sapiens Human MAGAASPCANGCGPGAPSDAEVLHLCRSLEVGTVMTLFYSKKSQRPERKTFQVKLETRQITWSRGADKIEGAIDIREIKEIRPGKTSRDFDRYQEDPAFRPDQSHCFVILYGMEFRLKTLSLQATSEDEVNMWIKGLTWLMEDTLQAPTPLQIERWLRKQFYSVDRNREDRISAKDLKNMLSQVNYRVPNMRFLRERLTDLEQRSGDITYGQFAQLYRSLMYSAQKTMDLPFLEASTLRAGERPELCRVSLPEFQQFLLDYQGELWAVDRLQVQEFMLSFLRDPLREIEEPYFFLDEFVTFLFSKENSVWNSQLDAVCPDTMNNPLSHYWISSSHNTYLTGDQFSSESSLEAYARCLRMGCRCIELDCWDGPDGMPVIYHGHTLTTKIKFSDVLHTIKEHAFVASEYPVILSIEDHCSIAQQRNMAQYFKKVLGDTLLTKPVEISADGLPSPNQLKRKILIKHKKLAEGSAYEEVPTSMMYSENDISNSIKNGILYLEDPVNHEWYPHYFVLTSSKIYYSEETSSDQGNEDEEEPKEVSSSTELHSNEKWFHGKLGAGRDGRHIAERLLTEYCIETGAPDGSFLVRESETFVGDYTLSFWRNGKVQHCRIHSRQDAGTPKFFLTDNLVFDSLYDLITHYQQVPLRCNEFEMRLSEPVPQTNAHESKEWYHASLTRAQAEHMLMRVPRDGAFLVRKRNEPNSYAISFRAEGKIKHCRVQQEGQTVMLGNSEFDSLVDLISYYEKHPLYRKMKLRYPINEEALEKIGTAEPDYGALYEGRNPGFYVEANPMPTFKCAVKALFDYKAQREDELTFIKSAIIQNVEKQEGGWWRGDYGGKKQLWFPSNYVEEMVNPVALEPEREHLDENSPLGDLLRGVLDVPACQIAIRPEGKNNRLFVFSISMASVAHWSLDVAADSQEELQDWVKKIREVAQTADARLTEGKIMERRKKIALELSELVVYCRPVPFDEEKIGTERACYRDMSSFPETKAEKYVNKAKGKKFLQYNRLQLSRIYPKGQRLDSSNYDPLPMWICGSQLVALNFQTPDKPMQMNQALFMTGRHCGYVLQPSTMRDEAFDPFDKSSLRGLEPCAISIEVLGARHLPKNGRGIVCPFVEIEVAGAEYDSTKQKTEFVVDNGLNPVWPAKPFHFQISNPEFAFLRFVVYEEDMFSDQNFLAQATFPVKGLKTGYRAVPLKNNYSEDLELASLLIKIDIFPAKENGDLSPFSGTSLRERGSDASGQLFHGRAREGSFESRYQQPFEDFRISQEHLADHFDSRERRAPRRTRVNGDNRL p39052-2 Dynamin-2 aa Dnm2 Dyn2 protein protein rat Rattus norvegicus (Rat) Rat MGNRGMEELIPLVNKLQDAFSSIGQSCHLDLPQIAVVGGQSAGKSSVLENFVGRDFLPRGSGIVTRRPLILQLIFSKTEYAEFLHCKSKKFTDFDEVRQEIEAETDRVTGTNKGISPVPINLRVYSPHVLNLTLIDLPGITKVPVGDQPPDIEYQIKDMILQFISRESSLILAVTPANMDLANSDALKLAKEVDPQGLRTIGVITKLDLMDEGTDARDVLENKLLPLRRGYIGVVNRSQKDIEGRKDIRAALAAERKFFLSHPAYRHMADRMGTPHLQKTLNQQLTNHIRESLPTLRSKLQSQLLSLEKEVEEYKNFRPDDPTRKTKALLQMVQQFGVDFEKRIEGSGDQVDTLELSGGARINRIFHERFPFELVKMEFDEKDLRREISYAIKNIHGVRTGLFTPDMAFEAIVKKQLVKLKEPSLKCVDLVVSELATVIKKCAEKLSSYPRLREETERIVTTYIREREGRTKDQILLLIDIEQSYINTNHEDFIGFANAQQRSTQLNKKRAIPNQGEILVIRRGWLTINNISLMKGGSKEYWFVLTAESLSWYKDEEEKEKKYMLPLDNLKIRDVEKGFMSNKHVFAIFNTEQRNVYKDLRQIELACDSQEDVDSWKASFLRAGVYPEKDQAENEDGAQENTFSMDPQLERQVETIRNLVDSYVAIINKSIRDLMPKTIMHLMINNTKAFIHHELLAYLYSSADQSSLMEESAEQAQRRDDMLRMYHALKEALNIIGDISTSTVSTPVPPPVDDTWLQNTSSHSPTPQRRPVSSVHPPGRPPAVRGPTPGPPLIPMPVGATSSFSAPPIPSRPGPQNVFANNDPFSAPPQIPSRPARIPPGIPPGVPSRRAPAAPSRPTIIRPAEPSLLD Sequestered to clathrin-coated pits and vesicles at the plasma membrane and the Golgi apparatus. cttn-dnm2 1314337 n/a 1314341 unspecified role unspecified role prey prey n/a 1314342 unspecified role unspecified role bait bait physical association physical association false false false Fig 1F (lane 3). The Dynamin coimmunoprecipitated with cortactin is confirmed to contain dynamin-2 by reblotting with a specific antibody. dnm2-cttn 1314337 n/a 1314341 unspecified role unspecified role bait bait n/a 1314342 unspecified role unspecified role prey prey physical association physical association false false false Fig 1g (lane 4). Cortactin was coimmunoprecipitated when Dynamin was immunoprecipitated with a Pan-dynamin antibody , but not when Dynamin was immunoprecipitated with a dynamin-2 tail-specific antibody. Dnm2 was confirmed to be present in the complex pulled down by the pan-dynamin antibody. cttn1-dnm2-1 1314338 n/a 1314343 unspecified role unspecified role bait bait sh3 domain sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 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 ecoli Escherichia coli (strain K12) n/a 1314341 unspecified role unspecified role prey prey mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells physical association physical association false false false Fig 2a (lane 2), 2b . The dymanin-cortactin interaction was inhibited in the presence of the poly-proline synthetic peptides P1 and P2 (containing the consensus motif PxxPSRP), but not in the presence of peptides P3, P4 and P5 (lacking the whole consensus motif). plcg1-dnm2-1 1314338 n/a 1314341 unspecified role unspecified role prey prey mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells n/a 1314344 unspecified role unspecified role bait bait sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 ecoli Escherichia coli (strain K12) physical association physical association false false false Fig 2C. PLCgamma-1 did not interact with dynamin-2 in the presence of peptides P1, P2 and p4. cttn1-dnm2-4 1314339 n/a 1314341 unspecified role unspecified role prey prey mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells n/a 1314343 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 sh3 domain sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig 3a. Dynamin interacted with cortactin in both PDGF-treated and -untreated calls. cttn-dnm2-5 1314340 n/a 1314345 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1314342 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig 4(d'' and e'').