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/ NKp44 triggers NK cell activation through DAP12 association that is not influenced by a putative cytoplasmic inhibitory sequence. NKp44 triggers NK cell activation through DAP12 association that is not influenced by a putative cytoplasmic inhibitory sequence. Journal of immunology (Baltimore, Md. : 1950) 2004 imex curation Campbell KS., Yusa S., Kikuchi-Maki A., Catina TL. Interactions using chimeric constructs have not been entered as they are of dubious biological relevance. Results from Y2H positive control interaction (CD5+ AP50) have not been included as the origin of AP50 could not be traced. Kerry.Campbell@fccc.edu 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions human-nk92 human NK cell line NK92 Human nk92 NK cell line NK92 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Campbell KS., Yusa S., Kikuchi-Maki A., Catina TL. Interactions using chimeric constructs have not been entered as they are of dubious biological relevance. Results from Y2H positive control interaction (CD5+ AP50) have not been included as the origin of AP50 could not be traced. I cannot enter fig 3C as an -Anti-Tag IP as no WT Flag-NKp44 is transfected - only mutant Kerry.Campbell@fccc.edu Accepted 2017-JUN-12 AT 08:11 BST AT 08:11 BST by ORCHARD Journal of immunology (Baltimore, Md. : 1950) 2004 imex curation I have added a Y2H interaction - not entered before as previous comment about AP50 origin - have now added as Human - see comment at experiment level about species of CD5. (CD5: species unclear; This paper stated "Mouse CD5", but via ref 15 -refers to "Raman 1998 (pmid:9668105)The entire 94-amino acid cytoplasmic domain of human CD5,...was fused to the GAL4 binding domain (BD) and used as a “bait” to screen an GAL4 activation domain (AD) cDNA library prepared from Epstein-Barr virus-transformed human peripheral blood lymphocytes. no mention of Mouse CD5. A BLAT of primers only gives Human.(mutations could be from either species)) NKp44 triggers NK cell activation through DAP12 association that is not influenced by a putative cytoplasmic inhibitory sequence. NKp44 triggers NK cell activation through DAP12 association that is not influenced by a putative cytoplasmic inhibitory sequence. Journal of immunology (Baltimore, Md. : 1950) 2004 imex curation Campbell KS., Yusa S., Kikuchi-Maki A., Catina TL. Interactions using chimeric constructs have not been entered as they are of dubious biological relevance. Results from Y2H positive control interaction (CD5+ AP50) have not been included as the origin of AP50 could not be traced. Kerry.Campbell@fccc.edu 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) 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 Campbell KS., Yusa S., Kikuchi-Maki A., Catina TL. Interactions using chimeric constructs have not been entered as they are of dubious biological relevance. Results from Y2H positive control interaction (CD5+ AP50) have not been included as the origin of AP50 could not be traced. AP50 has been assumed to be of Human origin as Ref 15 isolates AP50 from a human cDNA library. The species of CD5 is unclear; This paper methods section states "Mouse CD5 (aa 378–471)" provided by C. Raman ref 15. However this Ref 15 (Raman -1998 , pmid:9668105) states:- "The entire 94-amino acid cytoplasmic domain of human CD5,...was fused to the GAL4 binding domain (BD) and used as a “bait” to screen an GAL4 activation domain (AD) cDNA library prepared from Epstein-Barr virus-transformed human peripheral blood lymphocytes. A BLAT of primers only gives Human. Kerry.Campbell@fccc.edu Accepted 2017-JUN-12 AT 08:11 BST AT 08:11 BST by ORCHARD Journal of immunology (Baltimore, Md. : 1950) 2004 imex curation I have added a Y2H interaction - not entered before as previous comment about AP50 origin - have now added as Human - see comment at experiment level about species of CD5. (CD5: species unclear; This paper stated "Mouse CD5", but via ref 15 -refers to "Raman 1998 (pmid:9668105)The entire 94-amino acid cytoplasmic domain of human CD5,...was fused to the GAL4 binding domain (BD) and used as a “bait” to screen an GAL4 activation domain (AD) cDNA library prepared from Epstein-Barr virus-transformed human peripheral blood lymphocytes. no mention of Mouse CD5. A BLAT of primers only gives Human.(mutations could be from either species)) nctr2_human Natural cytotoxicity triggering receptor 2 NCR2 Lymphocyte antigen 95 homolog NK cell-activating receptor Natural killer cell p44-related protein LY95 protein protein human Homo sapiens Human MAWRALHPLLLLLLLFPGSQAQSKAQVLQSVAGQTLTVRCQYPPTGSLYEKKGWCKEASALVCIRLVTSSKPRTMAWTSRFTIWDDPDAGFFTVTMTDLREEDSGHYWCRIYRPSDNSVSKSVRFYLVVSPASASTQTSWTPRDLVSSQTQTQSCVPPTAGARQAPESPSTIPVPSQPQNSTLRPGPAAPIALVPVFCGLLVAKSLVLSALLVWWGDIWWKTMMELRSLDTQKATCHLQQVTDLPWTSVSSPVEREILYHTVARTKISDDDDEHTL 4B7AF3F451CA9F9E cd5_human T-cell surface glycoprotein CD5 CD5 LEU1 Lymphocyte antigen T1/Leu-1 protein protein human Homo sapiens Human MPMGSLQPLATLYLLGMLVASCLGRLSWYDPDFQARLTRSNSKCQGQLEVYLKDGWHMVCSQSWGRSSKQWEDPSQASKVCQRLNCGVPLSLGPFLVTYTPQSSIICYGQLGSFSNCSHSRNDMCHSLGLTCLEPQKTTPPTTRPPPTTTPEPTAPPRLQLVAQSGGQHCAGVVEFYSGSLGGTISYEAQDKTQDLENFLCNNLQCGSFLKHLPETEAGRAQDPGEPREHQPLPIQWKIQNSSCTSLEHCFRKIKPQKSGRVLALLCSGFQPKVQSRLVGGSSICEGTVEVRQGAQWAALCDSSSARSSLRWEEVCREQQCGSVNSYRVLDAGDPTSRGLFCPHQKLSQCHELWERNSYCKKVFVTCQDPNPAGLAAGTVASIILALVLLVVLLVVCGPLAYKKLVKKFRQKKQRQWIGPTGMNQNMSFHRNHTATVRSHAENPTASHVDNEYSQPPRNSHLSAYPALEGALHRSSMQPDNSSDSDYDLHGAQRL 9131AEC9683EE1D3 ap2m1_human AP-2 complex subunit mu Adaptor protein complex AP-2 subunit mu AP-2 mu chain AP2M1 CLAPM1 KIAA0109 Adaptin-mu2 Plasma membrane adaptor AP-2 50 kDa protein HA2 50 kDa subunit Clathrin coat assembly protein AP50 Clathrin coat-associated protein AP50 Clathrin assembly protein complex 2 mu medium chain Adaptor-related protein complex 2 subunit mu protein protein human Homo sapiens Human MIGGLFIYNHKGEVLISRVYRDDIGRNAVDAFRVNVIHARQQVRSPVTNIARTSFFHVKRSNIWLAAVTKQNVNAAMVFEFLYKMCDVMAAYFGKISEENIKNNFVLIYELLDEILDFGYPQNSETGALKTFITQQGIKSQHQTKEEQSQITSQVTGQIGWRREGIKYRRNELFLDVLESVNLLMSPQGQVLSAHVSGRVVMKSYLSGMPECKFGMNDKIVIEKQGKGTADETSKSGKQSIAIDDCTFHQCVRLSKFDSERSISFIPPDGEFELMRYRTTKDIILPFRVIPLVREVGRTKLEVKVVIKSNFKPSLLAQKIEVRIPTPLNTSGVQVICMKGKAKYKASENAIVWKIKRMAGMKESQISAEIELLPTNDKKKWARPPISMNFEVPFAPSGLKVRYLKVFEPKLNYSDHDVIKWVRYIGRSGIYETRC 82803219BA279954 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 ncr2-tyrobp-2 1 6 unspecified role unspecified role prey prey 3 unspecified role unspecified role bait bait physical association physical association figure 1, 3C dodecyl maltoside-detergent The presence of NKp44 with a K183A mutation resulted in a weaker DAP12 band. We confirmed that the mAb can immunoprecipitate NKp44 in association with DAP12, 2434-1 2 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 7G