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/ Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. BMC biology 2009 imex curation Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-neuron Mus musculus neuronal cell neuron Neuron [Neuronal cell; Neural cell anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. cortical neurons pengxiaozhong@pumc.edu.cn Accepted 2018-JUN-19 AT 12:56 BST AT 12:56 BST by MGT BMC biology 2009 Only protein-protein interactions imex curation imex curation sorry, I meant to combine Fig2A and Fig.3c; all done!; all done, new experiment created Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. BMC biology 2009 imex curation Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-brain-cortex Mus musculus brain cortex brain_cortex Brain cortex [Cerebral cortex; Cerebrum cortex; Cortex; Pallium] confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn Accepted 2018-JUN-19 AT 12:56 BST AT 12:56 BST by MGT BMC biology 2009 Only protein-protein interactions imex curation imex curation sorry, I meant to combine Fig2A and Fig.3c; all done!; all done, new experiment created Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. BMC biology 2009 imex curation Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae 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 Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn Accepted 2018-JUN-19 AT 12:56 BST AT 12:56 BST by MGT BMC biology 2009 Only protein-protein interactions imex curation imex curation sorry, I meant to combine Fig2A and Fig.3c; all done!; all done, new experiment created Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. BMC biology 2009 imex curation Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn Accepted 2018-JUN-19 AT 12:56 BST AT 12:56 BST by MGT BMC biology 2009 Only protein-protein interactions imex curation imex curation sorry, I meant to combine Fig2A and Fig.3c; all done!; all done, new experiment created Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. BMC biology 2009 imex curation Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn Accepted 2018-JUN-19 AT 12:56 BST AT 12:56 BST by MGT BMC biology 2009 Only protein-protein interactions imex curation imex curation sorry, I meant to combine Fig2A and Fig.3c; all done!; all done, new experiment created Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. Downstream of tyrosine kinase/docking protein 6, as a novel substrate of tropomyosin-related kinase C receptor, is involved in neurotrophin 3-mediated neurite outgrowth in mouse cortex neurons. BMC biology 2009 imex curation Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-brain Mus musculus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Li Wq., Shi L., You Yg., Gong Yh., Yin B., Yuan Jg., Peng Xz. pengxiaozhong@pumc.edu.cn Accepted 2018-JUN-19 AT 12:56 BST AT 12:56 BST by MGT BMC biology 2009 Only protein-protein interactions imex curation imex curation sorry, I meant to combine Fig2A and Fig.3c; all done!; all done, new experiment created dok6_human Docking protein 6 DOK6 DOK5L Downstream of tyrosine kinase 6 protein protein human Homo sapiens Human MASNFNDIVKQGYVKIRSRKLGIFRRCWLVFKKASSKGPRRLEKFPDEKAAYFRNFHKVTELHNIKNITRLPRETKKHAVAIIFHDETSKTFACESELEAEEWCKHLCMECLGTRLNDISLGEPDLLAAGVQREQNERFNVYLMPTPNLDIYGECTMQITHENIYLWDIHNAKVKLVMWPLSSLRRYGRDSTWFTFESGRMCDTGEGLFTFQTREGEMIYQKVHSATLAIAEQHERLMLEMEQKARLQTSLTEPMTLSKSISLPRSAYWHHITRQNSVGEIYSLQGHGFGSSKMSRAQTFPSYAPEQSEEAQQPLSRSSSYGFSYSSSLIQ 0C5EF2BF247091C1 ntrk3_human NT-3 growth factor receptor NTRK3 TRKC GP145-TrkC Neurotrophic tyrosine kinase receptor type 3 TrkC tyrosine kinase protein protein human Homo sapiens Human MDVSLCPAKCSFWRIFLLGSVWLDYVGSVLACPANCVCSKTEINCRRPDDGNLFPLLEGQDSGNSNGNASINITDISRNITSIHIENWRSLHTLNAVDMELYTGLQKLTIKNSGLRSIQPRAFAKNPHLRYINLSSNRLTTLSWQLFQTLSLRELQLEQNFFNCSCDIRWMQLWQEQGEAKLNSQNLYCINADGSQLPLFRMNISQCDLPEISVSHVNLTVREGDNAVITCNGSGSPLPDVDWIVTGLQSINTHQTNLNWTNVHAINLTLVNVTSEDNGFTLTCIAENVVGMSNASVALTVYYPPRVVSLEEPELRLEHCIEFVVRGNPPPTLHWLHNGQPLRESKIIHVEYYQEGEISEGCLLFNKPTHYNNGNYTLIAKNPLGTANQTINGHFLKEPFPESTDNFILFDEVSPTPPITVTHKPEEDTFGVSIAVGLAAFACVLLVVLFVMINKYGRRSKFGMKGPVAVISGEEDSASPLHHINHGITTPSSLDAGPDTVVIGMTRIPVIENPQYFRQGHNCHKPDTYVQHIKRRDIVLKRELGEGAFGKVFLAECYNLSPTKDKMLVAVKALKDPTLAARKDFQREAELLTNLQHEHIVKFYGVCGDGDPLIMVFEYMKHGDLNKFLRAHGPDAMILVDGQPRQAKGELGLSQMLHIASQIASGMVYLASQHFVHRDLATRNCLVGANLLVKIGDFGMSRDVYSTDYYRLFNPSGNDFCIWCEVGGHTMLPIRWMPPESIMYRKFTTESDVWSFGVILWEIFTYGKQPWFQLSNTEVIECITQGRVLERPRVCPKEVYDVMLGCWQREPQQRLNIKEIYKILHALGKATPIYLDILG DIP protein Q16288 original sequence version: 169 7FE8846830083C08 ntrk3_mouse NT-3 growth factor receptor Ntrk3 GP145-TrkC Neurotrophic tyrosine kinase receptor type 3 TrkC TrkC tyrosine kinase protein protein mouse Mus musculus Mouse MDVSLCPAKCSFWRIFLLGSVWLDYVGSVLACPANCVCSKTEINCRRPDDGNLFPLLEGQDSGNSNGNASINITDISRNITSIHIENWRGLHTLNAVDMELYTGLQKLTIKNSGLRNIQPRAFAKNPHLRYINLSSNRLTTLSWQLFQTLSLRELRLEQNFFNCSCDIRWMQLWQEQGEARLDSQSLYCISADGSQLPLFRMNISQCDLPEISVSHVNLTVREGDNAVITCNGSGSPLPDVDWIVTGLQSINTHQTNLNWTNVHAINLTLVNVTSEDNGFTLTCIAENVVGMSNASVALTVYYPPRVVSLVEPEVRLEHCIEFVVRGNPTPTLHWLYNGQPLRESKIIHMDYYQEGEVSEGCLLFNKPTHYNNGNYTLIAKNALGTANQTINGHFLKEPFPESTDFFDFESDASPTPPITVTHKPEEDTFGVSIAVGLAAFACVLLVVLFIMINKYGRRSKFGMKGPVAVISGEEDSASPLHHINHGITTPSSLDAGPDTVVIGMTRIPVIENPQYFRQGHNCHKPDTYVQHIKRRDIVLKRELGEGAFGKVFLAECYNLSPTKDKMLVAVKALKDPTLAARKDFQREAELLTNLQHEHIVKFYGVCGDGDPLIMVFEYMKHGDLNKFLRAHGPDAMILVDGQPRQAKGELGLSQMLHIASQIASGMVYLASQHFVHRDLATRNCLVGANLLVKIGDFGMSRDVYSTDYYRVGGHTMLPIRWMPPESIMYRKFTTESDVWSFGVILWEIFTYGKQPWFQLSNTEVIECITQGRVLERPRVCPKEVYDVMLGCWQREPQQRLNIKEIYKILHALGKATPIYLDILG 4DE08AB546CC5BD6 dok6_mouse Docking protein 6 Downstream of tyrosine kinase 6 Dok6 protein protein mouse Mus musculus Mouse MASNFNDIVKQGYVKIRSRKLGIFRRCWLVFKKASSKGPRRLEKFPDEKAAYFRNFHKVTELHNIKNITRLPRETKKHAVAIIFHDETSKTFACESELEAEEWCKHLCMECLGTRLNDISLGEPDLLAAGVQREQNERFNVYLMPTPNLDIYGECTMQITHENIYLWDIHNAKVKLVMWPLSSLRRYGRDSTWFTFESGRMCDTGEGLFTFQTREGEMIYQKVHSATLAIAEQHERLMLEMEQKARLQTSLTEPMTLSKSISLPRSAYWHHITRQNSVGEIYSLQGHGFGSSKMSRAQTFPSYAPEQSEEAQPPLSRSSSYGFSYSSSLIQ 0C5EF364547091C1 ntf3_human Neurotrophin-3 Neurotrophic factor HDNF Nerve growth factor 2 NTF3 protein protein human Homo sapiens Human MSILFYVIFLAYLRGIQGNNMDQRSLPEDSLNSLIIKLIQADILKNKLSKQMVDVKENYQSTLPKAEAPREPERGGPAKSAFQPVIAMDTELLRQQRRYNSPRVLLSDSTPLEPPPLYLMEDYVGSPVVANRTSRRKRYAEHKSHRGEYSVCDSESLWVTDKSSAIDIRGHQVTVLGEIKTGNSPVKQYFYETRCKEARPVKNGCRGIDDKHWNSQCKTSQTYVRALTSENNKLVGWRWIRIDTSCVCALSRKIGRT DIP protein P20783 original sequence version: 133 39A5BB3B28E25E03 dok6-trkc-1 3 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered cytosolic domain sufficient to bind sufficient binding region sufficient to bind range range range range Q16288:p.Lys572Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain K A Tue Jun 30 19:37:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys572Ala' Q16288:p.Tyr516Phe mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y F Tue Jun 30 17:52:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr516Phe' Q16288:p.Tyr821Phe mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain Y F Tue Jun 30 17:44:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr821Phe' Q16288:p.Tyr710Glu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Y E Tue Jun 30 20:16:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr710Glu' Q16288:p.Tyr709Asp mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Y D Tue Jun 30 15:35:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr709Asp' Q16288:p.Tyr705Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Y A Tue Jun 30 20:07:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr705Ala' 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 1C, Fig. 1D, Fig. 3B dok6-trkc-2 5 11 stimulator stimulator unspecified role unspecified role purified purified ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate engineered engineered Q16288:p.Lys572Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain K A Tue Jun 30 19:17:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys572Ala' Q16288:p.Tyr516Phe mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y F Tue Jun 30 17:02:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr516Phe' Q16288:p.Tyr821Phe mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Y F Tue Jun 30 17:34:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr821Phe' v5 tag v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 7 unspecified role unspecified role bait bait purified purified n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) association association Fig. 2A, Fig. 3C Consistent with results from the yeast two-hybrid assay findings, the precipitation results proved that the Dok6 PTB domain binds to the activated TrkC receptor strongly in vitro, while no remarkable binding was found between Dok6 and TrkA or TrkB with activation or no activation (Figure (Figure2a,2a, panel 1) Consistent with results from the yeast two-hybrid assay findings, the precipitation results proved that the Dok6 PTB domain binds to the activated TrkC receptor strongly in vitro, while no remarkable binding was found between Dok6 and TrkA or TrkB with activation or no activation (Figure (Figure2a,2a, panel 1) dok6-ntf3-1 4 11 stimulator stimulator unspecified role unspecified role purified purified ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered v5 tag v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered flag tag 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. 2B dok6-ntf3-2 6 9 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig. 4C dok6-ntrk3-1 2 9 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell 10 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization Fig. 4B Immunostaining showed that TrkC and Dok6 colocalise at the membrane, cytoplasm and neurites in primary cultured cortex neurons (Figure 4b) Immunostaining showed that TrkC and Dok6 colocalise at the membrane, cytoplasm and neurites in primary cultured cortex neurons (Figure 4b) dok6-ntrk3-2 1 11 stimulator stimulator unspecified role unspecified role purified purified 9 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association Fig. 6A Consistent with previous results, Dok6 binds to the TrkC receptor in cortex neurons upon NT-3 stimulation, while without NT-3 stimulation no obvious interactions exist (Figure (Figure6a).6a). Consistent with previous results, Dok6 binds to the TrkC receptor in cortex neurons upon NT-3 stimulation, while without NT-3 stimulation no obvious interactions exist (Figure 6a).