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/ Differential NKG2D binding to highly related human NKG2D ligands ULBP2 and RAET1G is determined by a single amino acid in the alpha2 domain. Differential NKG2D binding to highly related human NKG2D ligands ULBP2 and RAET1G is determined by a single amino acid in the alpha2 domain. European journal of immunology 2009 imex curation Wittenbrink M., Spreu J., Steinle A., Wittenbrink M., Spreu J., Steinle A. Results from figure 4 have not been annotated as chimeric proteins are used and the results are of dubious biological relevance. alexander.steinle@uni‐tuebingen.de 15-Nov-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro facs fluorescence-activated cell sorting FACS Flow cytometry facs tag fluorescence tag visualisation by fluorescence tag fluorescence Wittenbrink M., Spreu J., Steinle A., Wittenbrink M., Spreu J., Steinle A. Results from figure 4 have not been annotated as chimeric proteins are used and the results are of dubious biological relevance. I have made immobilised proteins as "Baits"- not usual for FACs. In some instances some (IgG1) Fc tagged proteins were attached to Protein A-coupled microspheres pre-loaded with anti-human IgG1 antibodies. alexander.steinle@uni‐tuebingen.de Accepted 2017-OCT-12 AT 11:25 BST AT 11:25 BST by ORCHARD European journal of immunology 2009 imex curation Differential NKG2D binding to highly related human NKG2D ligands ULBP2 and RAET1G is determined by a single amino acid in the alpha2 domain. Differential NKG2D binding to highly related human NKG2D ligands ULBP2 and RAET1G is determined by a single amino acid in the alpha2 domain. European journal of immunology 2009 imex curation Wittenbrink M., Spreu J., Steinle A., Wittenbrink M., Spreu J., Steinle A. Results from figure 4 have not been annotated as chimeric proteins are used and the results are of dubious biological relevance. alexander.steinle@uni‐tuebingen.de 15-Nov-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro facs fluorescence-activated cell sorting FACS Flow cytometry facs antibody detection identification by antibody antibody detection Wittenbrink M., Spreu J., Steinle A., Wittenbrink M., Spreu J., Steinle A. Results from figure 4 have not been annotated as chimeric proteins are used and the results are of dubious biological relevance. In some instances some Biotin tagged proteins were attached to microspheres pre-coupled to Streptavidin. alexander.steinle@uni‐tuebingen.de Accepted 2017-OCT-12 AT 11:25 BST AT 11:25 BST by ORCHARD European journal of immunology 2009 imex curation ul16p_hcmva Protein UL16 UL16 protein protein hcmva Human cytomegalovirus (strain AD169) (HHV-5) Human herpesvirus 5 HHV-5 MERRRGTVPLGWVFFVLCLSASSSCAVDLGSKSSNSTCRLNVTELASIHPGETWTLHGMCISICYYENVTEDEIIGVAFTWQHNESVVDLWLYQNDTVIRNFSDITTNILQDGLKMRTVPVTKLYTSRMVTNLTVGRYDCLRCENGTTKIIERLYVRLGSLYPRPPGSGLAKHPSVSADEELSATLARDIVLVSAITLFFFLLALRIPQRLCQRLRIRLPHRYQRLRTED B72F2C241C569967 ulbp5_human UL-16 binding protein 5 Retinoic acid early transcript 1G protein ULBP5 RAET1G protein protein human Homo sapiens Human MAAAASPAFLLRLPLLLLLSSWCRTGLADPHSLCYDITVIPKFRPGPRWCAVQGQVDEKTFLHYDCGSKTVTPVSPLGKKLNVTTAWKAQNPVLREVVDILTEQLLDIQLENYIPKEPLTLQARMSCEQKAEGHGSGSWQLSFDGQIFLLFDSENRMWTTVHPGARKMKEKWENDKDMTMSFHYISMGDCTGWLEDFLMGMDSTLEPSAGAPPTMSSGTAQPRATATTLILCCLLIMCLLICSRHSLTQSHGHHPQSLQPPPHPPLLHPTWLLRRVLWSDSYQIAKRPLSGGHVTRVTLPIIGDDSHSLPCPLALYTINNGAARYSEPLQVSIS 3F8350036951BA69 ulbp2_human UL16-binding protein 2 ULBP2 NKG2D ligand 2 N2DL2 RAET1H ALCAN-alpha Retinoic acid early transcript 1H UNQ463/PRO791 protein protein human Homo sapiens Human MAAAAATKILLCLPLLLLLSGWSRAGRADPHSLCYDITVIPKFRPGPRWCAVQGQVDEKTFLHYDCGNKTVTPVSPLGKKLNVTTAWKAQNPVLREVVDILTEQLRDIQLENYTPKEPLTLQARMSCEQKAEGHSSGSWQFSFDGQIFLLFDSEKRMWTTVHPGARKMKEKWENDKVVAMSFHYFSMGDCIGWLEDFLMGMDSTLEPSAGAPLAMSSGTTQLRATATTLILCCLLIILPCFILPGI 804D1A0D4A9C5CF8 nkg2d_human NKG2-D type II integral membrane protein KLRK1 NKG2D NKG2-D-activating NK receptor NK cell receptor D Killer cell lectin-like receptor subfamily K member 1 D12S2489E protein protein human Homo sapiens Human MGWIRGRRSRHSWEMSEFHNYNLDLKKSDFSTRWQKQRCPVVKSKCRENASPFFFCCFIAVAMGIRFIIMVAIWSAVFLNSLFNQEVQIPLTESYCGPCPKNWICYKNNCYQFFDESKNWYESQASCMSQNASLLKVYSKEDQDLLKLVKSYHWMGLVHIPTNGSWQWEDGSILSPNLLTIIEMQKGDCALYASSFKGYIENCSTPNTYICMQRTV DIP protein P26718 original sequence version: 106 C22F6BD533D7800E ul16-ulbp2-1 2 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n_terminus 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 chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 3 unspecified role unspecified role neutral component neutral component c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range signal+ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association 3A-2 klrk1-ulbp2-6 1 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed Q9BZM5:p.Phe141Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain F L Wed May 02 18:42:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe141leu' Tue Jun 30 20:07:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe141Leu' n_terminus 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 chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 6 unspecified role unspecified role neutral component neutral component c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range streptavidin-PE rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 18 19 physical association physical association 3A, Supp fig 3 I gave up trying to see if any construct existed with "Asn 80 through Thr 147" klrk1-ulbp2-4 1 6 unspecified role unspecified role prey prey c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range streptavidin-PE rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed signal+ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 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. 23 24 physical association physical association 3B weak binding ulbp2-klrk1-1 1 6 unspecified role unspecified role prey prey c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range streptavidin-PE rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc 31 32 physical association physical association 3B-2 strong binding ULBP2-Fc, which apparently interacted much stronger with NKG2D tetramers raet1g-klrk1-2 2 6 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecd sufficient to bind sufficient binding region sufficient to bind range range range range 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 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. physical association physical association 3C weak binding recombinant RAET1G-Fc (ULBP5) is NOT sold by R+D - so must be same construct as before. I can find no info on biotinylated-NKG2D that is NOT in a tetramer. klrk1-raet1g-1 2 6 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecd sufficient to bind sufficient binding region sufficient to bind range range range range 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc physical association physical association 3C-2 strong binding I can find no info on biotinylated-NKG2D that is NOT in a tetramer. klrk1-ulbp2-5 2 6 unspecified role unspecified role bait bait human-nkl Homo sapiens nkl nkl 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc physical association physical association 3D strong binding klrk1-raet1g-2 2 6 unspecified role unspecified role bait bait human-nkl Homo sapiens nkl nkl 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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. physical association physical association 3D weak binding recombinant RAET1G-Fc (ULBP5) is NOT sold by R+D - so must be same construct as before. klrk1-ulbp2-7 1 4 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed Q6H3X3:p.Leu141Phe mutation mutation certain certain sequence position certain certain certain sequence position certain L F Wed May 02 19:22:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu141phe' Tue Jun 30 20:05:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu141Phe' n_terminus 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 chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 6 unspecified role unspecified role neutral component neutral component c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range streptavidin-PE rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 68 67 physical association physical association Supp fig 3