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/ Ragulator-Rag complex targets mTORC1 to the lysosomal surface and is necessary for its activation by amino acids. Ragulator-Rag complex targets mTORC1 to the lysosomal surface and is necessary for its activation by amino acids. Cell (0092-8674) 2010 imex curation Sancak Y., Bar-Peled L., Zoncu R., Markhard AL., Nada S., Sabatini DM. Apoptosis - Interactions involving proteins with a function related to apoptosis I have not added colocalisation with lamp proteins as these are only organelle markers. Untagged proteins visualised by immunostaining with a fluorescently tagged secondary antibody. Source of RRAG family proteins taken from PMID: 18497260 as referenced in the text. Interactions involving Raptor, mTor, S6K1 and Rheb could not be annotated as the source of these proteins could not be traced. sabatini@wi.mit.edu 13-Sept-2010: Contacted by JYOTI. jyoti Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Sancak Y., Bar-Peled L., Zoncu R., Markhard AL., Nada S., Sabatini DM. Apoptosis - Interactions involving proteins with a function related to apoptosis I have not added colocalisation with lamp proteins as these are only organelle markers. Untagged proteins visualised by immunostaining with a fluorescently tagged secondary antibody. Source of RRAG family proteins taken from PMID: 18497260 as referenced in the text. Interactions involving Raptor, mTor, S6K1 and Rheb could not be annotated as the source of these proteins could not be traced. sabatini@wi.mit.edu Accepted 2010-JUL-30 by JYOTI. 13-Sept-2010: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions imex curation Ragulator-Rag complex targets mTORC1 to the lysosomal surface and is necessary for its activation by amino acids. Ragulator-Rag complex targets mTORC1 to the lysosomal surface and is necessary for its activation by amino acids. Cell (0092-8674) 2010 imex curation Sancak Y., Bar-Peled L., Zoncu R., Markhard AL., Nada S., Sabatini DM. Apoptosis - Interactions involving proteins with a function related to apoptosis I have not added colocalisation with lamp proteins as these are only organelle markers. Untagged proteins visualised by immunostaining with a fluorescently tagged secondary antibody. Source of RRAG family proteins taken from PMID: 18497260 as referenced in the text. Interactions involving Raptor, mTor, S6K1 and Rheb could not be annotated as the source of these proteins could not be traced. sabatini@wi.mit.edu 13-Sept-2010: Contacted by JYOTI. jyoti Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Sancak Y., Bar-Peled L., Zoncu R., Markhard AL., Nada S., Sabatini DM. Apoptosis - Interactions involving proteins with a function related to apoptosis I have not added colocalisation with lamp proteins as these are only organelle markers. Untagged proteins visualised by immunostaining with a fluorescently tagged secondary antibody. Source of RRAG family proteins taken from PMID: 18497260 as referenced in the text. Interactions involving Raptor, mTor, S6K1 and Rheb could not be annotated as the source of these proteins could not be traced. sabatini@wi.mit.edu Accepted 2010-JUL-30 by JYOTI. 13-Sept-2010: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions imex curation Ragulator-Rag complex targets mTORC1 to the lysosomal surface and is necessary for its activation by amino acids. Ragulator-Rag complex targets mTORC1 to the lysosomal surface and is necessary for its activation by amino acids. Cell (0092-8674) 2010 imex curation Sancak Y., Bar-Peled L., Zoncu R., Markhard AL., Nada S., Sabatini DM. Apoptosis - Interactions involving proteins with a function related to apoptosis I have not added colocalisation with lamp proteins as these are only organelle markers. Untagged proteins visualised by immunostaining with a fluorescently tagged secondary antibody. Source of RRAG family proteins taken from PMID: 18497260 as referenced in the text. Interactions involving Raptor, mTor, S6K1 and Rheb could not be annotated as the source of these proteins could not be traced. sabatini@wi.mit.edu 13-Sept-2010: Contacted by JYOTI. jyoti Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip sequence tag sequence tag identification sequence tag Sancak Y., Bar-Peled L., Zoncu R., Markhard AL., Nada S., Sabatini DM. Apoptosis - Interactions involving proteins with a function related to apoptosis I have not added colocalisation with lamp proteins as these are only organelle markers. Untagged proteins visualised by immunostaining with a fluorescently tagged secondary antibody. Source of RRAG family proteins taken from PMID: 18497260 as referenced in the text. Interactions involving Raptor, mTor, S6K1 and Rheb could not be annotated as the source of these proteins could not be traced. sabatini@wi.mit.edu Accepted 2010-JUL-30 by JYOTI. 13-Sept-2010: Contacted by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions imex curation Ragulator-Rag complex targets mTORC1 to the lysosomal surface and is necessary for its activation by amino acids. Ragulator-Rag complex targets mTORC1 to the lysosomal surface and is necessary for its activation by amino acids. Cell (0092-8674) 2010 imex curation Sancak Y., Bar-Peled L., Zoncu R., Markhard AL., Nada S., Sabatini DM. Apoptosis - Interactions involving proteins with a function related to apoptosis I have not added colocalisation with lamp proteins as these are only organelle markers. Untagged proteins visualised by immunostaining with a fluorescently tagged secondary antibody. Source of RRAG family proteins taken from PMID: 18497260 as referenced in the text. Interactions involving Raptor, mTor, S6K1 and Rheb could not be annotated as the source of these proteins could not be traced. sabatini@wi.mit.edu 13-Sept-2010: Contacted by JYOTI. jyoti Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Sancak Y., Bar-Peled L., Zoncu R., Markhard AL., Nada S., Sabatini DM. Apoptosis - Interactions involving proteins with a function related to apoptosis I have not added colocalisation with lamp proteins as these are only organelle markers. Untagged proteins visualised by immunostaining with a fluorescently tagged secondary antibody. Source of RRAG family proteins taken from PMID: 18497260 as referenced in the text. Interactions involving Raptor, mTor, S6K1 and Rheb could not be annotated as the source of these proteins could not be traced. sabatini@wi.mit.edu Accepted 2010-JUL-30 by JYOTI. Cell (0092-8674) 2010 jyoti Only protein-protein interactions imex curation ltor1_human Ragulator complex protein LAMTOR1 LAMTOR1 PP7157 p27Kip1-releasing factor from RhoA C11orf59 Lipid raft adaptor protein p18 Protein associated with DRMs and endosomes PDRO Late endosomal/lysosomal adaptor and MAPK and MTOR activator 1 protein protein human Homo sapiens Human MGCCYSSENEDSDQDREERKLLLDPSSPPTKALNGAEPNYHSLPSARTDEQALLSSILAKTASNIIDVSAADSQGMEQHEYMDRARQYSTRLAVLSSSLTHWKKLPPLPSLTSQPHQVLASEPIPFSDLQQVSRIAAYAYSALSQIRVDAKEELVVQFGIP DIP protein Q6IAA8 original sequence version: 88 610CC6C548356051 ltor2_human Ragulator complex protein LAMTOR2 LAMTOR2 MAPBPIP Late endosomal/lysosomal Mp1-interacting protein Roadblock domain-containing protein 3 Endosomal adaptor protein p14 HSPC003 ROBLD3 Late endosomal/lysosomal adaptor and MAPK and MTOR activator 2 Mitogen-activated protein-binding protein-interacting protein protein protein human Homo sapiens Human MLRPKALTQVLSQANTGGVQSTLLLNNEGSLLAYSGYGDTDARVTAAIASNIWAAYDRNGNQAFNEDNLKFILMDCMEGRVAITRVANLLLCMYAKETVGFGMLKAKAQALVQYLEEPLTQVAAS DIP protein Q9Y2Q5 original sequence version: 121 69599211C2CBC03D rraga_human Ras-related GTP-binding protein A Adenovirus E3 14.7 kDa-interacting protein 1 FIP-1 RRAGA protein protein human Homo sapiens Human MPNTAMKKKVLLMGKSGSGKTSMRSIIFANYIARDTRRLGATIDVEHSHVRFLGNLVLNLWDCGGQDTFMENYFTSQRDNIFRNVEVLIYVFDVESRELEKDMHYYQSCLEAILQNSPDAKIFCLVHKMDLVQEDQRDLIFKEREEDLRRLSRPLECACFRTSIWDETLYKAWSSIVYQLIPNVQQLEMNLRNFAQIIEADEVLLFERATFLVISHYQCKEQRDVHRFEKISNIIKQFKLSCSKLAASFQSMEVRNSNFAAFIDIFTSNTYVMVVMSDPSIPSAATLINIRNARKHFEKLERVDGPKHSLLMR DIP protein Q7L523 original sequence version: 116 DIP protein Q7L523 original sequence version: 104 B0DA1FC8FA6B766A rragd_human Ras-related GTP-binding protein D RRAGD protein protein human Homo sapiens Human MSQVLGKPQPQDEDDAEEEEEEDELVGLADYGDGPDSSDADPDSGTEEGVLDFSDPFSTEVKPRILLMGLRRSGKSSIQKVVFHKMSPNETLFLESTNKICREDVSNSSFVNFQIWDFPGQIDFFDPTFDYEMIFRGTGALIFVIDSQDDYMEALARLHLTVTRAYKVNTDINFEVFIHKVDGLSDDHKIETQRDIHQRANDDLADAGLEKIHLSFYLTSIYDHSIFEAFSKVVQKLIPQLPTLENLLNIFISNSGIEKAFLFDVVSKIYIATDSTPVDMQTYELCCDMIDVVIDISCIYGLKEDGAGTPYDKESTAIIKLNNTTVLYLKEVTKFLALVCFVREESFERKGLIDYNFHCFRKAIHEVFEVRMKVVKSRKVQNRLQKKKRATPNGTPRVLL 96FF0854B11AA1BC rragb_human Ras-related GTP-binding protein B RRAGB protein protein human Homo sapiens Human MEESDSEKTTEKENLGPRMDPPLGEPEGSLGWVLPNTAMKKKVLLMGKSGSGKTSMRSIIFANYIARDTRRLGATILDRIHSLQINSSLSTYSLVDSVGNTKTFDVEHSHVRFLGNLVLNLWDCGGQDTFMENYFTSQRDNIFRNVEVLIYVFDVESRELEKDMHYYQSCLEAILQNSPDAKIFCLVHKMDLVQEDQRDLIFKEREEDLRRLSRPLECSCFRTSIWDETLYKAWSSIVYQLIPNVQQLEMNLRNFAEIIEADEVLLFERATFLVISHYQCKEQRDAHRFEKISNIIKQFKLSCSKLAASFQSMEVRNSNFAAFIDIFTSNTYVMVVMSDPSIPSAATLINIRNARKHFEKLERVDGPKQCLLMR DIP protein Q5VZM2 original sequence version: 89 0D32F7D86E2A00DE ltor3_human Ragulator complex protein LAMTOR3 MAP2K1IP1 Mitogen-activated protein kinase kinase 1-interacting protein 1 LAMTOR3 PRO2783 MEK-binding partner 1 MAPKSP1 Late endosomal/lysosomal adaptor and MAPK and MTOR activator 3 Mitogen-activated protein kinase scaffold protein 1 protein protein human Homo sapiens Human MADDLKRFLYKKLPSVEGLHAIVVSDRDGVPVIKVANDNAPEHALRPGFLSTFALATDQGSKLGLSKNKSIICYYNTYQVVQFNRLPLVVSFIASSSANTGLIVSLEKELAPLFEELRQVVEVS 0A5196CF41D83D13 rragc_human Ras-related GTP-binding protein C GTPase-interacting protein 2 TIB929 RRAGC protein protein human Homo sapiens Human MSLQYGAEETPLAGSYGAADSFPKDFGYGVEEEEEEAAAAGGGVGAGAGGGCGPGGADSSKPRILLMGLRRSGKSSIQKVVFHKMSPNETLFLESTNKIYKDDISNSSFVNFQIWDFPGQMDFFDPTFDYEMIFRGTGALIYVIDAQDDYMEALTRLHITVSKAYKVNPDMNFEVFIHKVDGLSDDHKIETQRDIHQRANDDLADAGLEKLHLSFYLTSIYDHSIFEAFSKVVQKLIPQLPTLENLLNIFISNSGIEKAFLFDVVSKIYIATDSSPVDMQSYELCCDMIDVVIDVSCIYGLKEDGSGSAYDKESMAIIKLNNTTVLYLKEVTKFLALVCILREESFERKGLIDYNFHCFRKAIHEVFEVGVTSHRSCGHQTSASSLKALTHNGTPRNAI DIP protein Q9HB90 original sequence version: 120 DIP protein Q9HB90 original sequence version: 109 35B3171253CC69CF ragc-raga-1 2 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association RagC interacts with RagA and RagB independently of amino acid availability. Fig. 1D rragb-robld3-1 3 9 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association Mass spectrometric analysis of anti-FLAG immunoprecipitates prepared from human HEK293T cells stably expressing FLAG-RagB or FLAG-RagD, but not FLAG-Rap2a, consistently revealed the presence of proteins encoded by the MAPKSP1, ROBLD3, and c11orf59 genes (Figure 2A). Fig. 2A rragd-robld3-1 3 8 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association Mass spectrometric analysis of anti-FLAG immunoprecipitates prepared from human HEK293T cells stably expressing FLAG-RagB or FLAG-RagD, but not FLAG-Rap2a, consistently revealed the presence of proteins encoded by the MAPKSP1, ROBLD3, and c11orf59 genes (Figure 2A). Fig. 2A robld3-mpks1-1 1 9 unspecified role unspecified role prey prey 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 region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey 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 region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 2B robld3-mpks1-2 1 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 2D rragb-mapksp1-1 1 9 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 region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey association association Fig. 2E Recombinant RagB-RagD heterodimers coimmunoprecipitate endogenous p14, MP1, and p18. rragb-mapksp1-2 2 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait association association Fig. 2F Endogenous RagC coimmunoprecipitates endogenous p14 and MP1. rragb-mapksp1-3 1 9 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association Fig. 2I Endogenous Ragulator coimmunoprecipitates with FLAG-RagB independently of amino acid availability and GTP-loading of RagB. ragc-c11orf59-1 4 11 unspecified role unspecified role neutral component neutral component 5 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 colocalization colocalization Fig. 2J