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/ A split-luciferase complementation, real-time reporting assay enables monitoring of the disease-associated transmembrane protein TREM2 in live cells. A split-luciferase complementation, real-time reporting assay enables monitoring of the disease-associated transmembrane protein TREM2 in live cells. The Journal of biological chemistry 2017 imex curation Varnum MM., Clayton KA., Yoshii-Kitahara A., Yonemoto G., Koro L., Ikezu S., Ikezu T. Alzheimers - Interactions investigated in the context of Alzheimers disease tikezu@bu.edu 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. fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Varnum MM., Clayton KA., Yoshii-Kitahara A., Yonemoto G., Koro L., Ikezu S., Ikezu T. Alzheimers - Interactions investigated in the context of Alzheimers disease tikezu@bu.edu Accepted 2019-MAR-04 AT 16:41 GMT AT 16:41 GMT by LIVIAP The Journal of biological chemistry 2017 Only protein-protein interactions imex curation imex curation A split-luciferase complementation, real-time reporting assay enables monitoring of the disease-associated transmembrane protein TREM2 in live cells. A split-luciferase complementation, real-time reporting assay enables monitoring of the disease-associated transmembrane protein TREM2 in live cells. The Journal of biological chemistry 2017 imex curation Varnum MM., Clayton KA., Yoshii-Kitahara A., Yonemoto G., Koro L., Ikezu S., Ikezu T. Alzheimers - Interactions investigated in the context of Alzheimers disease tikezu@bu.edu 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. split renilla luciferase complementation Split renilla luciferase complementation renilla luciferase renilla luciferase tag luciferase tag visualisation by luciferase assay tag luciferase Varnum MM., Clayton KA., Yoshii-Kitahara A., Yonemoto G., Koro L., Ikezu S., Ikezu T. Alzheimers - Interactions investigated in the context of Alzheimers disease tikezu@bu.edu Accepted 2019-MAR-04 AT 16:41 GMT AT 16:41 GMT by LIVIAP The Journal of biological chemistry 2017 Only protein-protein interactions imex curation imex curation A split-luciferase complementation, real-time reporting assay enables monitoring of the disease-associated transmembrane protein TREM2 in live cells. A split-luciferase complementation, real-time reporting assay enables monitoring of the disease-associated transmembrane protein TREM2 in live cells. The Journal of biological chemistry 2017 imex curation Varnum MM., Clayton KA., Yoshii-Kitahara A., Yonemoto G., Koro L., Ikezu S., Ikezu T. Alzheimers - Interactions investigated in the context of Alzheimers disease tikezu@bu.edu 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. crosslink cross-linking study crosslink antibody detection identification by antibody antibody detection Varnum MM., Clayton KA., Yoshii-Kitahara A., Yonemoto G., Koro L., Ikezu S., Ikezu T. Alzheimers - Interactions investigated in the context of Alzheimers disease Identification of the TREM2-TYROBP interaction was by sandwich ELISA. Cells were treated with the tyrosine phosphatase inhibitor sodium orthovanadate and cross-linker dimethyl 3,3′-dithiobispropionimidate (DTBP). tikezu@bu.edu Accepted 2019-MAR-04 AT 16:41 GMT AT 16:41 GMT by LIVIAP The Journal of biological chemistry 2017 Only protein-protein interactions imex curation imex curation tyobp_human TYRO protein tyrosine kinase-binding protein TYROBP DAP12 KARAP DNAX-activation protein 12 Killer-activating receptor-associated protein protein protein human Homo sapiens Human MGGLEPCSRLLLLPLLLAVSGLRPVQAQAQSDCSCSTVSPGVLAGIVMGDLVLTVLIALAVYFLGRLVPRGRGAAEAATRKQRITETESPYQELQGQRSDVYSDLNTQRPYYK 267CB1C1756F89F0 trem2_human Triggering receptor expressed on myeloid cells 2 TREM2 Triggering receptor expressed on monocytes 2 protein protein human Homo sapiens Human MEPLRLLILLFVTELSGAHNTTVFQGVAGQSLQVSCPYDSMKHWGRRKAWCRQLGEKGPCQRVVSTHNLWLLSFLRRWNGSTAITDDTLGGTLTITLRNLQPHDAGLYQCQSLHGSEADTLRKVLVEVLADPLDHRDAGDLWFPGESESFEDAHVEHSISRSLLEGEIPFPPTSILLLLACIFLIKILAASALWAAAWHGQKPGTHPPSELDCGHDPGYQLQTLPGLRDT C894AA210F708AF7 trem2-tyrobp-1 2 4 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed c-terminal renilla-n renilla-n c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed Q9NZC2:p.Thr66Met mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain T M Tue Jun 30 18:44:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr66Met' c-terminal renilla-c renilla-c c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q9NZC2:p.Arg47His 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 R H Tue Jun 30 16:23:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg47His' Q9NZC2:p.Ser116Cys 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 S C Tue Jun 30 16:21:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser116Cys' physical association physical association For examination of the real-time protein-protein interaction between TREM2 and TYROBP, we developed a mammalian expression vector (TREM2-CLuc-IRES-TYROBP-NLuc) that utilizes the “bait-prey” split-Renilla luciferase complementation assay (Fig. 1A) For examination of the real-time protein-protein interaction between TREM2 and TYROBP, we developed a mammalian expression vector (TREM2-CLuc-IRES-TYROBP-NLuc) that utilizes the “bait-prey” split-Renilla luciferase complementation assay (Fig. 1A) Fig. 2, 6 Supp. Fig. 1 TREM2-cross-linked antibody The TREM2 the Thr66Met mutation did not show enhanced luciferase activity on anti-TREM2 antibody stimulation trem2-tyrobp-2 3 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed c-terminal renilla-n renilla-n c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminal renilla-c renilla-c c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig. 3 TREM2-cross-linked antibody trem2-tyrobp-3 1 4 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed c-terminal renilla-n renilla-n c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed Q9NZC2:p.Ser116Cys 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 S C Tue Jun 30 20:04:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser116Cys' Q9NZC2:p.Thr66Met 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 T M Tue Jun 30 19:08:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr66Met' c-terminal renilla-c renilla-c c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q9NZC2:p.Arg47His 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 R H Tue Jun 30 20:16:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg47His' colocalization colocalization Immunostaining of permeabilized cells showed mostly similar distribution of TREM2 and TYROBP across all groups, indicative of their intracellular interactions (supplemental Fig. S3A). Supp. Fig. 3A