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 immunoreceptor TIGIT regulates antitumor and antiviral CD8(+) T cell effector function. The immunoreceptor TIGIT regulates antitumor and antiviral CD8(+) T cell effector function. Cancer cell 2014 imex curation Johnston RJ., Comps-Agrar L., Hackney J., Yu X., Huseni M., Yang Y., Park S., Javinal V., Chiu H., Irving B., Eaton DL., Grogan JL. Cancer - Interactions investigated in the context of cancer grogan.jane@gene.com Only protein-protein interactions imex curation human-t-cell Homo sapiens t-cell Peripheral blood T-lymphocytes fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret antibody detection identification by antibody antibody detection Johnston RJ., Comps-Agrar L., Hackney J., Yu X., Huseni M., Yang Y., Park S., Javinal V., Chiu H., Irving B., Eaton DL., Grogan JL. Cancer - Interactions investigated in the context of cancer After stimulation with anti-CD3 and Cd28 antibodies, T cells were sorted on the basis of TIGIT expression. Endogenous TIGIT and CD226 were labelled with donor flurophore-conjugated anti-TIGIT and acceptor fluorophore-conjugated anti-CD226 antibodies. grogan.jane@gene.com Accepted 2015-JUN-17 by ORCHARD Cancer cell 2014 Only protein-protein interactions imex curation imex curation The immunoreceptor TIGIT regulates antitumor and antiviral CD8(+) T cell effector function. The immunoreceptor TIGIT regulates antitumor and antiviral CD8(+) T cell effector function. Cancer cell 2014 imex curation Johnston RJ., Comps-Agrar L., Hackney J., Yu X., Huseni M., Yang Y., Park S., Javinal V., Chiu H., Irving B., Eaton DL., Grogan JL. Cancer - Interactions investigated in the context of cancer grogan.jane@gene.com Only protein-protein interactions imex curation in vitro In vitro fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Johnston RJ., Comps-Agrar L., Hackney J., Yu X., Huseni M., Yang Y., Park S., Javinal V., Chiu H., Irving B., Eaton DL., Grogan JL. Cancer - Interactions investigated in the context of cancer I have added "antibody detection" to the TIGIT molecule for Interaction detection at Feature level. Figure 7D:-Cells were transfected with N-terminus SNAP-tagged (ST) CD226. The SNAP tags were then labelled with either donor-conjugated benzylguanine SNAP-Lumi-4Tb or acceptor-conjugated benzyl-guanine SNAP-A647. Figure 7F:- Cells were transfected with N-terminus SNAP-tagged (ST) CD226 or HA-TIGIT. The SNAP tag was then labelled with acceptor conjugated benzyl-guanine SNAP-A647 and the HA tag was labelled with donor-conjugated anti-HA Lumi-4Tb. grogan.jane@gene.com Accepted 2015-JUN-17 by ORCHARD Cancer cell 2014 Only protein-protein interactions imex curation imex curation I really dont know if we should add "donor/acceptor roles when on the Abs"- I also added a role of "fluorescence donor" to the TIGIT mol in Expt-1 at Participant level, when detected by antibody;- the other mol was tagged- so if I can (had to)add it in that instance, why not here also? It is FRET - and we usually add flu donor/acceptor for this Interaction Det method. (this is a rather unusual example anyway - unlikely to be repeated?) The immunoreceptor TIGIT regulates antitumor and antiviral CD8(+) T cell effector function. The immunoreceptor TIGIT regulates antitumor and antiviral CD8(+) T cell effector function. Cancer cell 2014 imex curation Johnston RJ., Comps-Agrar L., Hackney J., Yu X., Huseni M., Yang Y., Park S., Javinal V., Chiu H., Irving B., Eaton DL., Grogan JL. Cancer - Interactions investigated in the context of cancer grogan.jane@gene.com Only protein-protein interactions imex curation chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Johnston RJ., Comps-Agrar L., Hackney J., Yu X., Huseni M., Yang Y., Park S., Javinal V., Chiu H., Irving B., Eaton DL., Grogan JL. Cancer - Interactions investigated in the context of cancer grogan.jane@gene.com Accepted 2015-JUN-17 by ORCHARD Cancer cell 2014 Only protein-protein interactions imex curation imex curation I really dont know if we should add "donor/acceptor roles when on the Abs"- I also added a role of "fluorescence donor" to the TIGIT mol in Expt-1 at Participant level, when detected by antibody;- the other mol was tagged- so if I can (had to)add it in that instance, why not here also? It is FRET - and we usually add flu donor/acceptor for this Interaction Det method. (this is a rather unusual example anyway - unlikely to be repeated?) tigit_human T-cell immunoreceptor with Ig and ITIM domains TIGIT VSIG9 VSTM3 V-set and immunoglobulin domain-containing protein 9 V-set and transmembrane domain-containing protein 3 protein protein human Homo sapiens Human MRWCLLLIWAQGLRQAPLASGMMTGTIETTGNISAEKGGSIILQCHLSSTTAQVTQVNWEQQDQLLAICNADLGWHISPSFKDRVAPGPGLGLTLQSLTVNDTGEYFCIYHTYPDGTYTGRIFLEVLESSVAEHGARFQIPLLGAMAATLVVICTAVIVVVALTRKKKALRIHSVEGDLRRKSAGQEEWSPSAPSPPGSCVQAEAAPAGLCGEQRGEDCAELHDYFNVLSYRSLGNCSFFTETG B890AF73F618C851 cd226_human CD226 antigen CD226 DNAM1 DNAX accessory molecule 1 protein protein human Homo sapiens Human MDYPTLLLALLHVYRALCEEVLWHTSVPFAENMSLECVYPSMGILTQVEWFKIGTQQDSIAIFSPTHGMVIRKPYAERVYFLNSTMASNNMTLFFRNASEDDVGYYSCSLYTYPQGTWQKVIQVVQSDSFEAAVPSNSHIVSEPGKNVTLTCQPQMTWPVQAVRWEKIQPRQIDLLTYCNLVHGRNFTSKFPRQIVSNCSHGRWSVIVIPDVTVSDSGLYRCYLQASAGENETFVMRLTVAEGKTDNQYTLFVAGGTVLLLLFVISITTIIVIFLNRRRRRERRDLFTESWDTQKAPNNYRSPISTSQPTNQSMDDTREDIYVNYPTFSRRPKTRV 5379182CC4695097 cd226-1 2 5 unspecified role unspecified role fluorescence donor fluorescence donor N/A fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range N/A 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 crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. 5 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept N/A fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range N/A 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 crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. physical association physical association 7 D/E HA-TIGIT (T-cell immunoreceptor with Ig and ITIM domains) tigit-cd226-2 3 4 unspecified role unspecified role prey prey N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role bait bait N/A flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 7G cd226-tigit-1 2 4 antibody detection identification by antibody antibody detection tag fluorescence tag visualisation by fluorescence tag fluorescence unspecified role unspecified role fluorescence donor fluorescence donor N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. 5 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept N/A 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 N/A fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. physical association physical association 7 F/H anti-TIGIT antibody tigit-cd226-1 1 5 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept N/A flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role fluorescence donor fluorescence donor N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range proximity proximity close-contact colocalization neighbourhood interaction 7 I cd226-tigit-2 3 4 unspecified role unspecified role bait bait N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role prey prey N/A flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 7Gi