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/ Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Biochemistry (0006-2960) 1999 Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. n.j.gay@bioc.cam.ac.uk davet yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. n.j.gay@bioc.cam.ac.uk Accepted 2007-OCT-18 by DAVET. Biochemistry (0006-2960) 1999 davet Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Biochemistry (0006-2960) 1999 Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. n.j.gay@bioc.cam.ac.uk davet in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. n.j.gay@bioc.cam.ac.uk Accepted 2007-OCT-18 by DAVET. Biochemistry (0006-2960) 1999 davet Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Biochemistry (0006-2960) 1999 Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. n.j.gay@bioc.cam.ac.uk davet in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. Column of Sephacryl S200 used. n.j.gay@bioc.cam.ac.uk Accepted 2007-OCT-18 by DAVET. Biochemistry (0006-2960) 1999 davet Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Biochemistry (0006-2960) 1999 Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. n.j.gay@bioc.cam.ac.uk davet in vitro In vitro cosedimentation cosedimentation predetermined predetermined participant predetermined Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. n.j.gay@bioc.cam.ac.uk Accepted 2007-OCT-18 by DAVET. Biochemistry (0006-2960) 1999 davet Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Formation and biochemical characterization of tube/pelle death domain complexes: critical regulators of postreceptor signaling by the Drosophila toll receptor. Biochemistry (0006-2960) 1999 Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. n.j.gay@bioc.cam.ac.uk davet in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Schiffmann DA., White JH., Cooper A., Nutley MA., Harding SE., Jumel K., Solari R., Ray KP., Gay NJ. n.j.gay@bioc.cam.ac.uk Accepted 2007-OCT-18 by DAVET. Biochemistry (0006-2960) 1999 davet kpel_drome Serine/threonine-protein kinase pelle CG5974 pll protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MSGVQTAEAEAQAQNQANGNRTRSRSHLDNTMAIRLLPLPVRAQLCAHLDALDVWQQLATAVKLYPDQVEQISSQKQRGRSASNEFLNIWGGQYNHTVQTLFALFKKLKLHNAMRLIKDYVSEDLHKYIPRSVPTISELRAAPDSSAKVNNGPPFPSSSGVSNSNNNRTSTTATEEIPSLESLGNIHISTVQRAAESLLEIDYAELENATDGWSPDNRLGQGGFGDVYRGKWKQLDVAIKVMNYRSPNIDQKMVELQQSYNELKYLNSIRHDNILALYGYSIKGGKPCLVYQLMKGGSLEARLRAHKAQNPLPALTWQQRFSISLGTARGIYFLHTARGTPLIHGDIKPANILLDQCLQPKIGDFGLVREGPKSLDAVVEVNKVFGTKIYLPPEFRNFRQLSTGVDVYSFGIVLLEVFTGRQVTDRVPENETKKNLLDYVKQQWRQNRMELLEKHLAAPMGKELDMCMCAIEAGLHCTALDPQDRPSMNAVLKRFEPFVTD 4B29E2B40ACB81A8 tube_drome Protein Tube CG10520 tub tube protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MAYGWNGCGMGVQVNGSNGAIGLSSKYSRNTELRRVEDNDIYRLAKILDENSCWRKLMSIIPKGMDVQACSGAGCLNFPAEIKKGFKYTAQDVFQIDEAANRLPPDQSKSQMMIDEWKTSGKLNERPTVGVLLQLLVQAELFSAADFVALDFLNESTPARPVDGPGALISLELLEEEMEVDNEGLSLKYQSSTATLGADAQGSVGLNLDNFEKDIVRRDKSVPQPSGNTPPIAPPRRQQRSTTNSNFATLTGTGTTSTTIPNVPNLTILNPSEQIQEPVLQPRPMNIPDLSILISNSGDLRATVSDNPSNRTSSTDPPNIPRITLLIDNSGDVNSRPNHAPAKASTATTSTASSNNLPMISALNISKGSRETLRPESRSSSSSLSKDDDDDNDGEEDGEEEYPDAFLPNLSNSEQQSSNNDSSLTTVTGTSGDNSFELTNDSSSTSNDDYACNIPDLSELQQ DDFBB9D72B8036FC tub-pell 1 7 unspecified role unspecified role prey prey n terminal sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1393527]The shifted ranges (1-257) couldn't be applied as the new full feature sequence is different from the previous one. [EBI-1393527]1-258 [EBI-1393527]P22812,3 6 unspecified role unspecified role bait bait n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 2 Results of two-hybrid experiments (Figure 2 and Table 1) indicated that the N-tube but not the tube C-terminal fusion protein was capable of interacting with N-pelle. tub-pell-1 3 7 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1393541]The shifted ranges (1-257) couldn't be applied as the new full feature sequence is different from the previous one. [EBI-1393541]1-258 [EBI-1393541]P22812,3 6 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction 3 the protein complex elutes from the column in a single peak (lanes 8-12) separated from the slight excess of the N-tube monomer (lanes 15-16). This shows that the two proteins form a complex on mixing, most likely a heterodimer tub-pell-2 5 6 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1393560]The shifted ranges (1-257) couldn't be applied as the new full feature sequence is different from the previous one. [EBI-1393560]1-258 [EBI-1393560]P22812,3 direct interaction direct interaction 5 Analysis of the Tube/Pelle Interaction Using Surface Plasmon Resonance (Biacore) Measurements. tub-pell-3 4 6 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1393574]The shifted ranges (1-257) couldn't be applied as the new full feature sequence is different from the previous one. [EBI-1393574]1-258 [EBI-1393574]P22812,3 direct interaction direct interaction 6 the N-tube/N-pelle mixture was considered as a 1:1 heteromeric complex, evidence for which comes from the sedimentation equilibrium experiments tub-pell-4 2 7 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1393591]The shifted ranges (1-257) couldn't be applied as the new full feature sequence is different from the previous one. [EBI-1393591]1-258 [EBI-1393591]P22812,3 6 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction 7 the interaction between N-tube and N-pelle was characterized using isothermal titration microcalorimetry (ITC)