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/ The endosomal protein Appl1 mediates Akt substrate specificity and cell survival in vertebrate development. The endosomal protein Appl1 mediates Akt substrate specificity and cell survival in vertebrate development. Cell (0092-8674) 2008 imex curation Schenck A., Goto-Silva L., Collinet C., Rhinn M., Giner A., Habermann B., Brand M., Zerial M. The interactions involving AKT could not be curated As it is not clear which isoforms were detected. The source of the GSK-3beta construct could not be ascertained and thus interactions involving this construct could not be curated. zerial@mpi-cbg.de 27-Aug-2008: Contacted by JYOTI. davet Only protein-protein interactions 3 in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Schenck A., Goto-Silva L., Collinet C., Rhinn M., Giner A., Habermann B., Brand M., Zerial M. The interactions involving AKT could not be curated As it is not clear which isoforms were detected. The source of the GSK-3beta construct could not be ascertained and thus interactions involving this construct could not be curated. zerial@mpi-cbg.de Accepted 2008-SEP-6 by DAVET. Cell (0092-8674) 2008 davet Only protein-protein interactions 3 imex curation The endosomal protein Appl1 mediates Akt substrate specificity and cell survival in vertebrate development. The endosomal protein Appl1 mediates Akt substrate specificity and cell survival in vertebrate development. Cell (0092-8674) 2008 imex curation Schenck A., Goto-Silva L., Collinet C., Rhinn M., Giner A., Habermann B., Brand M., Zerial M. The interactions involving AKT could not be curated As it is not clear which isoforms were detected. The source of the GSK-3beta construct could not be ascertained and thus interactions involving this construct could not be curated. zerial@mpi-cbg.de 27-Aug-2008: Contacted by JYOTI. davet Only protein-protein interactions 3 danre-embryo Danio rerio Embryo Brachydanio rerio embryo Embryo Defined as the development stage prior to the initiation of tissue differentiation. confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Schenck A., Goto-Silva L., Collinet C., Rhinn M., Giner A., Habermann B., Brand M., Zerial M. Zebrafish embryos injected with 100 pg appl1-Venus mRNA at single cell stage and with 50 pg CFP-Rab5C mRNA into single blastomeres at 16-cell stage, fixed, and imaged at 50% epiboly stage. The interactions involving AKT could not be curated As it is not clear which isoforms were detected. The source of the GSK-3beta construct could not be ascertained and thus interactions involving this construct could not be curated. zerial@mpi-cbg.de Accepted 2008-SEP-6 by DAVET. 27-Aug-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 3 imex curation b2xcm0_danre appl2 protein protein danre Danio rerio (Zebrafish) Brachydanio rerio Zebrafish MPAVHHKLLLEEALQDSPQTRSLLSVFEEDAGTLTNYTNQLLQSMQRVFGAQNEMALATEQLSKQLLEYEKQNFALAKGDEEVINTLQQFAKSVEELNALHSELSTQIADKMVFPMVQFREKDLTEISTLKEIFSIATDEHEAAMVKYSRLPKKRENEKVKAEVVREVAYSRRKQHQAAMQYYCALNALQYRKRVAMLEPMLGYTHAQINFFKKGMELVSKKLDSFLNSVSNMNESIESQLEIDAEVMRSSQRELLSVDDSVYMPDHDQAAVSRALIQKAGYLNIRNKTGLVTTAWDRLFFFTQGGNLMCQPRGAVAGGMVLDLDNSSVMAVDCEDRRYCFQITSPNGKTSLILQAESKREYEEWICTLNNISRQIYLTDNPEAVAIKLHQTALQAVTPITSFEKRAEGSPNPDRAKPGGLPVTSQEAEKSSVSLEPEDLIVPGTPIQFDIMLPASEFLDQNRAGARRTNPFGETEEDDDESDTHDSLLQQVFAVRFLGSMAVRSGHTQEVIYEAMRQVLAARAIHNIFKTTESHLMISSSCIRLIDPQTQVTKISFQLQEVSEFAAHQENERLMGFVVRSSAEDDEPSFSVFVFESNTEGEKICYTINLAKEIADARKDPEALAQLMKSMPLTNDGKFLLLDNETGDTAETAGQEQQESEA 07B9AE411E8468C8 b2xcl9_danre appl1 protein protein danre Danio rerio (Zebrafish) Brachydanio rerio Zebrafish MPGIDKLPIEETLEDSPQTRSLLGVFEEDAAAISSYFSQLFKAMQRIYDAQNELSAATHLTSKLLKDYEKQRFPLGGDDEVMSSTLQQFAKVIDELSSCHAVLSTQLADAMMFPITQFQERDLREILILKEVFQLASDDHDAAVHRYSRLSKRKENEKVKNEVMEDVYTSRKKQHETTMHYFASLNMLQYKKKIALLEPLLGYMQAQISFFKLGSENLTQQWEDFLTNIGTSVQNVRREMDSDAEAMEQTIQDLKEASDPLYMPDPDPNRIPVNRNLTRKAGYLNTRNKTGLVSSSWERQYFFTQGGNLMSQSRGAVAGGLVMDIDNCSVMAVDCEDRRFCFQITAFDGKKAAVLQADSRKDCEEWIATINNISKRIYLSENPEEIAARVNQSALEAVTPSPSFQQRHESFRPNTQGKVKASRSGSQCSVGSESPALSSLSLDSLVAPDTPIQFDIISPVSEEHTSQTKSSAQGRTNPFGESGGSQSEESEDSILHQLFIVRFLGSMEVKATESSDVIYETMRQILAARAIHNIFRMTESHLLVTCECLKLIDPQTQVTRLRFPLSSVVLCASHQENKRLFGFVLQTAGGRADGQPVTVCYIFESNNDGEKICDSIGLAKQIALHSEMDRKASQKQRELDKAKEKQQEELHKQKQIEKDLEEQSRLIAASSRPNQPAADGQFLVLSNSQSEDSDVGEEGLKKGESEA B60136D26BE6C962 q7zvp2_danre rab5c protein protein danre Danio rerio (Zebrafish) Brachydanio rerio Zebrafish MAGRGGPARTNGAAPVGNKICQFKLVLLGESAVGKSSLVLRFVKGQFHEYQESTIGAAFLTQTLCLDDTTVKFEIWDTAGQERYHSLAPMYYRGAQAAIVVYDITNTDTFTRAKNWVKELQRQASPNIVIALAGNKADLANKRAVDFQEAQAYADDNSLLFMETSAKTAMNVNEIFMAIAKKLPKSEPQGGAGSGGRARGGVDLQETAPQGRSGQCCGGGN 019B355683BF71FA rab5c-appl1 1 5 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 ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction 1 D, S5 the ability of fish Appl proteins to bind Rab5 was tested in a GST pull-down assay using recombinant GST-Rab5C preloaded with either GTPgS or GDP and in vitro translated Appl1 and Appl2. They strongly and specifically bound to the active form of Rab5C EBI-1778249 gene EU053152 This interaction was stimulated when RAB5C was preloaded with GTP as compared to GDP. rab5c-appl1-1 2 5 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role neutral component neutral component region Venus (variant of YFP) yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization 1 B, C, S2 EBI-1778249; Q7ZVP2 injection EBI-1778249 gene EU053152 In coexpressing cells, Appl1- and Appl2-Venus colocalized with CFP-Rab5C in a characteristic endosomal pattern rab5c-appl2 1 3 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 5 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction 1 D the ability of fish Appl proteins to bind Rab5 was tested in a GST pull-down assay using recombinant GST-Rab5C preloaded with either GTPgS or GDP and in vitro translated Appl1 and Appl2. They strongly and specifically bound to the active form of Rab5C EBI-1778258 gene EU053153 This interaction was stimulated when RAB5C was preloaded with GTP as compared to GDP.