InnateDB InnateDB call-2010-2 The structural basis for intramembrane assembly of an activating immunoreceptor complex. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Call ME., Wucherpfennig KW., Chou JJ. resolved on non-denaturing gel, all proteins were in-vitro translated and audiolabelled chou@cmcd.hms.harvard.edu Nat. Immunol. (1529-2908) 2010 Only protein-protein interactions imex curation Curated by: InnateDB - a knowledge resource for innate immunity call-2010-1 The structural basis for intramembrane assembly of an activating immunoreceptor complex. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Call ME., Wucherpfennig KW., Chou JJ. chou@cmcd.hms.harvard.edu Nat. Immunol. (1529-2908) 2010 Only protein-protein interactions imex curation Curated by: InnateDB - a knowledge resource for innate immunity call-2010-4 The structural basis for intramembrane assembly of an activating immunoreceptor complex. in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Call ME., Wucherpfennig KW., Chou JJ. resolved on non-denaturing gel, all proteins were in-vitro translated and audiolabelled chou@cmcd.hms.harvard.edu Nat. Immunol. (1529-2908) 2010 Only protein-protein interactions imex curation Curated by: InnateDB - a knowledge resource for innate immunity call-2010-3 The structural basis for intramembrane assembly of an activating immunoreceptor complex. in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Call ME., Wucherpfennig KW., Chou JJ. resolved on non-denaturing gel, all proteins were in-vitro translated and audiolabelled chou@cmcd.hms.harvard.edu Nat. Immunol. (1529-2908) 2010 Only protein-protein interactions imex curation Curated by: InnateDB - a knowledge resource for innate immunity nkg2c_human NKG2-C type II integral membrane protein KLRC2 NKG2C CD159 antigen-like family member C NK cell receptor C NKG2-C-activating NK receptor protein protein human Homo sapiens Human MSKQRGTFSEVSLAQDPKRQQRKPKGNKSSISGTEQEIFQVELNLQNPSLNHQGIDKIYDCQGLLPPPEKLTAEVLGIICIVLMATVLKTIVLIPFLEQNNSSPNTRTQKARHCGHCPEEWITYSNSCYYIGKERRTWEESLLACTSKNSSLLSIDNEEEMKFLASILPSSWIGVFRNSSHHPWVTINGLAFKHKIKDSDNAELNCAVLQVNRLKSAQCGSSMIYHCKHKL 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 ki2s3_human Killer cell immunoglobulin-like receptor 2DS3 KIR2DS3 NKAT7 MHC class I NK cell receptor Natural killer-associated transcript 7 protein protein human Homo sapiens Human MSLMVISMACVGFFWLQGAWPHEGFRRKPSLLAHPGRLVKSEETVILQCWSDVMFEHFLLHREGTFNDTLRLIGEHIDGVSKANFSIGRMRQDLAGTYRCYGSVPHSPYQFSAPSDPLDIVITGLYEKPSLSAQPGPTVLAGESVTLSCSSWSSYDMYHLSTEGEAHERRFSAGPKVNGTFQADFPLGPATQGGTYRCFGSFHDSPYEWSKSSDPLLVSVTGNPSNSWPSPTEPSSKTGNPRHLHVLIGTSVVKLPFTILLFFLLHRWCSDKKNASVMDQGPAGNRTVNREDSDEQDHQEVSYA 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 hcst_human Hematopoietic cell signal transducer DAP10 KAP10 PIK3AP DNAX-activation protein 10 Membrane protein DAP10 Transmembrane adapter protein KAP10 UNQ587/PRO1157 HCST protein protein human Homo sapiens Human MIHLGHILFLLLLPVAAAQTTPGERSSLPAFYPGTSGSCSGCGSLSLPLLAGLVAADAVASLLIVGAVFLCARPRRSPAQEDGKVYINMPGRG dap12-tm_nkg2c-tm_fusion_protein DAP12tm-NKG2Ctm protein protein human Homo sapiens Human CSTVSPGVLAGIVVGDLVLTVLIALAVYFLGRLGTAEVLGIISIVLVATVLKTIVLIPFLEQN cd3d_human T-cell surface glycoprotein CD3 delta chain CD3D T3D T-cell receptor T3 delta chain protein protein human Homo sapiens Human MEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGIIVTDVIATLLLALGVFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNK cd3e_human T-cell surface glycoprotein CD3 epsilon chain CD3E T3E T-cell surface antigen T3/Leu-4 epsilon chain protein protein human Homo sapiens Human MQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI cd3g_human T-cell surface glycoprotein CD3 gamma chain CD3G T3G T-cell receptor T3 gamma chain protein protein human Homo sapiens Human MEQGKGLAVLILAIILLQGTLAQSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATISGFLFAEIVSIFVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQGNQLRRN tyrobp-tyrobp 1131581 n/a 1131585 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1131586 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false O43914:p.Thr54Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false O43914:p.Ala58Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 physical association physical association false false false Fig. 2B both DAP12 monomer and dimer were detected in the non-denaturing gel tyrobp-tyrobp-1 1131581 n/a 1131587 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1131586 unspecified role unspecified role bait bait O43914:p.Ala58Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false O43914:p.Thr54Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 physical association physical association false false false Fig. 2C both DAP12 monomer and dimer were detected in the non-denaturing gel hcst-hcst 1131581 n/a 1131588 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1131589 unspecified role unspecified role bait bait Q9UBK5:p.Asp57Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Stoichiometry: 2.0 physical association physical association false false false Fig 5D native gel on left showed oxidized NKG2D as a defused band, +DTT treatment (right) was performed for quantification of NKG2D proteins. tyrobp 1131582 n/a 1131586 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 2h label 2h label deuterium D2 2H2 2H2 deuterium D2 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 13c label 13c label C13 13C C13 13C undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 2D, 2E, 3A, 3C, S2C, S3, Table 1. klrc2-tyrobp-1 1131582 n/a 1131590 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 2h label 2h label deuterium D2 2H2 2H2 deuterium D2 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 13c label 13c label C13 13C C13 13C undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1131586 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 2h label 2h label deuterium D2 2H2 2H2 deuterium D2 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 13c label 13c label C13 13C C13 13C undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Fig. 2D, 2E, 3A, 3C, S2C, S3, Table 1. A pure disulfide-linked TYROBP transmembrane(TM)-TYROBP(TM)-KLRC2(TM) trimeric peptide product was used in the study. cd3e-cd3d 1131583 n/a 1131591 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1131592 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Fig. 5E TCRa was also co-immunoprecipitated in this experiment but was not included as a participant since the specific cDNA isoform/variable region could not be determined from the article nor from author correspondence cd3e-cd3g 1131583 n/a 1131593 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1131592 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Fig. 5F TCRb was also co-immunoprecipitated in this experiment but was not included as a participant since the specific cDNA isoform/variable region could not be determined from the article nor from author correspondence tyrobp-tyrobp-3 1131584 n/a 1131586 unspecified role unspecified role neutral component neutral component O43914:p.[Gly41Phe;Gly45Phe;Gly49Phe] mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O43914:p.[Asp50Ala;Thr54Ala] mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false O43914:p.Leu16Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false O43914:p.Ile46_Val47delinsAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 2F