InnateDB InnateDB The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils. The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils. Nat. Immunol. (1529-2908) 2012 imex curation Honda F., Kano H., Kanegane H., Nonoyama S., Kim E.S., Lee S.K., Takagi M., Mizutani S., Morio T. Apoptosis - Interactions involving proteins with a function related to apoptosis neutrophils were from healthy controls and patients with X-linked agammaglobulinemia (Btk deficient) tmorio.ped@tmd.ac.jp 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions Curated by: InnateDB - a knowledge resource for innate immunity human-neutrophil Homo sapiens polymorphonuclear neutrophil neutrophil Neutrophil [Neutrophilic granulocyte] pull down pull down western blot western blot Immuno blot Immuno blot Honda F., Kano H., Kanegane H., Nonoyama S., Kim E.S., Lee S.K., Takagi M., Mizutani S., Morio T. Apoptosis - Interactions involving proteins with a function related to apoptosis neutrophils were from healthy controls and patients with X-linked agammaglobulinemia (Btk deficient) tmorio.ped@tmd.ac.jp Accepted 2012-AUG-06 by MGT 19-Sept-2012: Contacted by IntAct-Help. cell source Phorbol 12-myristate 13-acetate (ng/ml) Nat. Immunol. (1529-2908) 2012 Only protein-protein interactions imex curation Curated by: InnateDB - a knowledge resource for innate immunity The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils. The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils. Nat. Immunol. (1529-2908) 2012 imex curation Honda F., Kano H., Kanegane H., Nonoyama S., Kim E.S., Lee S.K., Takagi M., Mizutani S., Morio T. Apoptosis - Interactions involving proteins with a function related to apoptosis neutrophils were from healthy controls and patients with X-linked agammaglobulinemia (Btk deficient) tmorio.ped@tmd.ac.jp 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions Curated by: InnateDB - a knowledge resource for innate immunity human-neutrophil Homo sapiens polymorphonuclear neutrophil neutrophil Neutrophil [Neutrophilic granulocyte] confocal microscopy confocal microscopy immunostaining immunostaining Honda F., Kano H., Kanegane H., Nonoyama S., Kim E.S., Lee S.K., Takagi M., Mizutani S., Morio T. Apoptosis - Interactions involving proteins with a function related to apoptosis neutrophils were from healthy controls and patients with X-linked agammaglobulinemia (Btk deficient) tmorio.ped@tmd.ac.jp Accepted 2012-AUG-06 by MGT 19-Sept-2012: Contacted by IntAct-Help. cell source Phorbol 12-myristate 13-acetate (ng/ml) Nat. Immunol. (1529-2908) 2012 Only protein-protein interactions imex curation Curated by: InnateDB - a knowledge resource for innate immunity The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils. The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils. Nat. Immunol. (1529-2908) 2012 imex curation Honda F., Kano H., Kanegane H., Nonoyama S., Kim E.S., Lee S.K., Takagi M., Mizutani S., Morio T. Apoptosis - Interactions involving proteins with a function related to apoptosis neutrophils were from healthy controls and patients with X-linked agammaglobulinemia (Btk deficient) tmorio.ped@tmd.ac.jp 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions Curated by: InnateDB - a knowledge resource for innate immunity human-neutrophil Homo sapiens polymorphonuclear neutrophil neutrophil Neutrophil [Neutrophilic granulocyte] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Honda F., Kano H., Kanegane H., Nonoyama S., Kim E.S., Lee S.K., Takagi M., Mizutani S., Morio T. Apoptosis - Interactions involving proteins with a function related to apoptosis neutrophils were from healthy controls and patients with X-linked agammaglobulinemia (Btk deficient) tmorio.ped@tmd.ac.jp Accepted 2012-AUG-06 by MGT 19-Sept-2012: Contacted by IntAct-Help. cell source Phorbol 12-myristate 13-acetate (ng/ml) Nat. Immunol. (1529-2908) 2012 Only protein-protein interactions imex curation Curated by: InnateDB - a knowledge resource for innate immunity cy24b_human Cytochrome b-245 heavy chain CYBB NOX2 CGD91-phox Cytochrome b(558) subunit beta Heme-binding membrane glycoprotein gp91phox NADPH oxidase 2 Neutrophil cytochrome b 91 kDa polypeptide Superoxide-generating NADPH oxidase heavy chain subunit gp91-1 gp91-phox p22 phagocyte B-cytochrome protein protein human Homo sapiens Human MGNWAVNEGLSIFVILVWLGLNVFLFVWYYRVYDIPPKFFYTRKLLGSALALARAPAACLNFNCMLILLPVCRNLLSFLRGSSACCSTRVRRQLDRNLTFHKMVAWMIALHSAIHTIAHLFNVEWCVNARVNNSDPYSVALSELGDRQNESYLNFARKRIKNPEGGLYLAVTLLAGITGVVITLCLILIITSSTKTIRRSYFEVFWYTHHLFVIFFIGLAIHGAERIVRGQTAESLAVHNITVCEQKISEWGKIKECPIPQFAGNPPMTWKWIVGPMFLYLCERLVRFWRSQQKVVITKVVTHPFKTIELQMKKKGFKMEVGQYIFVKCPKVSKLEWHPFTLTSAPEEDFFSIHIRIVGDWTEGLFNACGCDKQEFQDAWKLPKIAVDGPFGTASEDVFSYEVVMLVGAGIGVTPFASILKSVWYKYCNNATNLKLKKIYFYWLCRDTHAFEWFADLLQLLESQMQERNNAGFLSYNIYLTGWDESQANHFAVHHDEEKDVITGLKQKTLYGRPNWDNEFKTIASQHPNTRIGVFLCGPEALAETLSKQSISNSESGPRGVHFIFNKENF DIP protein P04839 original sequence version: 111 7E84051BD4000CE3 ksyk_human Tyrosine-protein kinase SYK SYK Spleen tyrosine kinase p72-Syk protein protein human Homo sapiens Human MASSGMADSANHLPFFFGNITREEAEDYLVQGGMSDGLYLLRQSRNYLGGFALSVAHGRKAHHYTIERELNGTYAIAGGRTHASPADLCHYHSQESDGLVCLLKKPFNRPQGVQPKTGPFEDLKENLIREYVKQTWNLQGQALEQAIISQKPQLEKLIATTAHEKMPWFHGKISREESEQIVLIGSKTNGKFLIRARDNNGSYALCLLHEGKVLHYRIDKDKTGKLSIPEGKKFDTLWQLVEHYSYKADGLLRVLTVPCQKIGTQGNVNFGGRPQLPGSHPATWSAGGIISRIKSYSFPKPGHRKSSPAQGNRQESTVSFNPYEPELAPWAADKGPQREALPMDTEVYESPYADPEEIRPKEVYLDRKLLTLEDKELGSGNFGTVKKGYYQMKKVVKTVAVKILKNEANDPALKDELLAEANVMQQLDNPYIVRMIGICEAESWMLVMEMAELGPLNKYLQQNRHVKDKNIIELVHQVSMGMKYLEESNFVHRDLAARNVLLVTQHYAKISDFGLSKALRADENYYKAQTHGKWPVKWYAPECINYYKFSSKSDVWSFGVLMWEAFSYGQKPYRGMKGSEVTAMLEKGERMGCPAGCPREMYDLMNLCWTYDVENRPGFAAVELRLRNYYYDVVN DIP protein P43405 original sequence version: 146 EAA6BDE65881FC68 lyn_human Tyrosine-protein kinase Lyn JTK8 Lck/Yes-related novel protein tyrosine kinase V-yes-1 Yamaguchi sarcoma viral related oncogene homolog p53Lyn p56Lyn LYN protein protein human Homo sapiens Human MGCIKSKGKDSLSDDGVDLKTQPVRNTERTIYVRDPTSNKQQRPVPESQLLPGQRFQTKDPEEQGDIVVALYPYDGIHPDDLSFKKGEKMKVLEEHGEWWKAKSLLTKKEGFIPSNYVAKLNTLETEEWFFKDITRKDAERQLLAPGNSAGAFLIRESETLKGSFSLSVRDFDPVHGDVIKHYKIRSLDNGGYYISPRITFPCISDMIKHYQKQADGLCRRLEKACISPKPQKPWDKDAWEIPRESIKLVKRLGAGQFGEVWMGYYNNSTKVAVKTLKPGTMSVQAFLEEANLMKTLQHDKLVRLYAVVTREEPIYIITEYMAKGSLLDFLKSDEGGKVLLPKLIDFSAQIAEGMAYIERKNYIHRDLRAANVLVSESLMCKIADFGLARVIEDNEYTAREGAKFPIKWTAPEAINFGCFTIKSDVWSFGILLYEIVTYGKIPYPGRTNADVMTALSQGYRMPRVENCPDELYDIMKMCWKEKAEERPTFDYLQSVLDDFYTATEGQYQQQP DIP protein P07948 original sequence version: 141 408D3D461204E378 src_human Proto-oncogene tyrosine-protein kinase Src pp60c-src SRC SRC1 Proto-oncogene c-Src protein protein human Homo sapiens Human MGSNKSKPKDASQRRRSLEPAENVHGAGGGAFPASQTPSKPASADGHRGPSAAFAPAAAEPKLFGGFNSSDTVTSPQRAGPLAGGVTTFVALYDYESRTETDLSFKKGERLQIVNNTEGDWWLAHSLSTGQTGYIPSNYVAPSDSIQAEEWYFGKITRRESERLLLNAENPRGTFLVRESETTKGAYCLSVSDFDNAKGLNVKHYKIRKLDSGGFYITSRTQFNSLQQLVAYYSKHADGLCHRLTTVCPTSKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTRVAIKTLKPGTMSPEAFLQEAQVMKKLRHEKLVQLYAVVSEEPIYIVTEYMSKGSLLDFLKGETGKYLRLPQLVDMAAQIASGMAYVERMNYVHRDLRAANILVGENLVCKVADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELTTKGRVPYPGMVNREVLDQVERGYRMPCPPECPESLHDLMCQCWRKEPEERPTFEYLQAFLEDYFTSTEPQYQPGENL DIP protein P12931 original sequence version: 188 DIP protein P12931 original sequence version: 189 DIP protein P12931 original sequence version: 190 DIP protein P12931 original sequence version: 191 DIP protein P12931 original sequence version: 115 DIP protein P12931 original sequence version: 116 DIP protein P12931 original sequence version: 164 DIP protein P12931 original sequence version: 142 DIP protein P12931 original sequence version: 143 DIP protein P12931 original sequence version: 150 DIP protein P12931 original sequence version: 160 DIP protein P12931 original sequence version: 184 DIP protein P12931 original sequence version: 155 DIP protein P12931-1 original sequence version: 155 DIP protein P12931 original sequence version: 202 C1908084683E5DE8 btk_human Tyrosine-protein kinase BTK Bruton tyrosine kinase B-cell progenitor kinase BPK BTK AGMX1 ATK Agammaglobulinemia tyrosine kinase protein protein human Homo sapiens Human MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES DIP protein Q06187 original sequence version: 118 DIP protein Q06187 original sequence version: 143 DF06B5D1FEC257CC mal_human Myelin and lymphocyte protein MAL T-lymphocyte maturation-associated protein protein protein human Homo sapiens Human MAPAAATGGSTLPSGFSVFTTLPDLLFIFEFIFGGLVWILVASSLVPWPLVQGWVMFVSVFCFVATTTLIILYIIGAHGGETSWVTLDAAYHCTAALFYLSASVLEALATITMQDGFTYRHYHENIAAVVFSYIATLLYVVHAVFSLIRWKSS F821FC1C0ADBE775 p85a_human Phosphatidylinositol 3-kinase regulatory subunit alpha PIK3R1 GRB1 Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha protein protein human Homo sapiens Human MSAEGYQYRALYDYKKEREEDIDLHLGDILTVNKGSLVALGFSDGQEARPEEIGWLNGYNETTGERGDFPGTYVEYIGRKKISPPTPKPRPPRPLPVAPGSSKTEADVEQQALTLPDLAEQFAPPDIAPPLLIKLVEAIEKKGLECSTLYRTQSSSNLAELRQLLDCDTPSVDLEMIDVHVLADAFKRYLLDLPNPVIPAAVYSEMISLAPEVQSSEEYIQLLKKLIRSPSIPHQYWLTLQYLLKHFFKLSQTSSKNLLNARVLSEIFSPMLFRFSAASSDNTENLIKVIEILISTEWNERQPAPALPPKPPKPTTVANNGMNNNMSLQDAEWYWGDISREEVNEKLRDTADGTFLVRDASTKMHGDYTLTLRKGGNNKLIKIFHRDGKYGFSDPLTFSSVVELINHYRNESLAQYNPKLDVKLLYPVSKYQQDQVVKEDNIEAVGKKLHEYNTQFQEKSREYDRLYEEYTRTSQEIQMKRTAIEAFNETIKIFEEQCQTQERYSKEYIEKFKREGNEKEIQRIMHNYDKLKSRISEIIDSRRRLEEDLKKQAAEYREIDKRMNSIKPDLIQLRKTRDQYLMWLTQKGVRQKKLNEWLGNENTEDQYSLVEDDEDLPHHDEKTWNVGSSNRNKAENLLRGKRDGTFLVRESSKQGCYACSVVVDGEVKHCVINKTATGYGFAEPYNLYSSLKELVLHYQHTSLVQHNDSLNVTLAYPVYAQQRR DIP protein P27986 original sequence version: 181 DIP protein P27986 original sequence version: 184 DIP protein P27986-1 original sequence version: 135 DIP protein P27986 original sequence version: 135 DIP protein P27986-1 original sequence version: 145 DIP protein P27986-1 original sequence version: 147 DIP protein P27986 original sequence version: 191 B9DAD8416C33140F rac2_human Ras-related C3 botulinum toxin substrate 2 p21-Rac2 Small G protein GX RAC2 protein protein human Homo sapiens Human MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVMVDSKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASYENVRAKWFPEVRHHCPSTPIILVGTKLDLRDDKDTIEKLKEKKLAPITYPQGLALAKEIDSVKYLECSALTQRGLKTVFDEAIRAVLCPQPTRQQKRACSLL DIP protein P15153 original sequence version: 154 DIP protein P15153 original sequence version: 153 DIP protein P15153 original sequence version: 123 2A1F1266B07C3210 fak1_human Focal adhesion kinase 1 PTK2 FAK FAK1 pp125FAK Protein-tyrosine kinase 2 Focal adhesion kinase-related nonkinase Protein phosphatase 1 regulatory subunit 71 p125FAK protein protein human Homo sapiens Human MAAAYLDPNLNHTPNSSTKTHLGTGMERSPGAMERVLKVFHYFESNSEPTTWASIIRHGDATDVRGIIQKIVDSHKVKHVACYGFRLSHLRSEEVHWLHVDMGVSSVREKYELAHPPEEWKYELRIRYLPKGFLNQFTEDKPTLNFFYQQVKSDYMLEIADQVDQEIALKLGCLEIRRSYWEMRGNALEKKSNYEVLEKDVGLKRFFPKSLLDSVKAKTLRKLIQQTFRQFANLNREESILKFFEILSPVYRFDKECFKCALGSSWIISVELAIGPEEGISYLTDKGCNPTHLADFTQVQTIQYSNSEDKDRKGMLQLKIAGAPEPLTVTAPSLTIAENMADLIDGYCRLVNGTSQSFIIRPQKEGERALPSIPKLANSEKQGMRTHAVSVSETDDYAEIIDEEDTYTMPSTRDYEIQRERIELGRCIGEGQFGDVHQGIYMSPENPALAVAIKTCKNCTSDSVREKFLQEALTMRQFDHPHIVKLIGVITENPVWIIMELCTLGELRSFLQVRKYSLDLASLILYAYQLSTALAYLESKRFVHRDIAARNVLVSSNDCVKLGDFGLSRYMEDSTYYKASKGKLPIKWMAPESINFRRFTSASDVWMFGVCMWEILMHGVKPFQGVKNNDVIGRIENGERLPMPPNCPPTLYSLMTKCWAYDPSRRPRFTELKAQLSTILEEEKAQQEERMRMESRRQATVSWDSGGSDEAPPKPSRPGYPSPRSSEGFYPSPQHMVQTNHYQVSGYPGSHGITAMAGSIYPGQASLLDQTDSWNHRPQEIAMWQPNVEDSTVLDLRGIGQVLPTHLMEERLIRQQQEMEEDQRWLEKEERFLKPDVRLSRGSIDREDGSLQGPIGNQHIYQPVGKPDPAAPPKKPPRPGAPGHLGSLASLSSPADSYNEGVKLQPQEISPPPTANLDRSNDKVYENVTGLVKAVIEMSSKIQPAPPEEYVPMVKEVGLALRTLLATVDETIPLLPASTHREIEMAQKLLNSDLGELINKMKLAQQYVMTSLQQEYKKQMLTAAHALAVDAKNLLDVIDQARLKMLGQTRPH D8A4C15138AB0243 cybb-rac2 2 4 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 4E. Co-localization of gp91 and Rac2 was observed in the membranes of resting Btk-deficient neutrophils but not in neutrophils from healthy controls. XLA cells 0 btk-mal 2 9 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization 0 Fig. 6C. Healthy cells mal-pik3r1 2 9 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component colocalization colocalization 100 (15 mins) Fig. 6E. Healthy cells mal-src 2 7 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 8D. Healthy/control cells c-Src is associated with Mal in the cytoplasm but not in the membrane of normal resting neutrophils. 0 btk-mal-1 3 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association Fig. 6A. PMA 0 ng/ml Healthy cells This interaction was not observed in healthy cells after PMA stimulation. mal-btk 3 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Fig. 6B. Healthy control cells PMA - 0 ng/ml pik3r1-mal 3 10 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association Fig. 6D. XLA (Btk deficient) neutrophils. Interaction detected only in XLA neutrophils. mal-ptk2 3 12 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association Fig. S6. Interaction detected only in XLA neutrophils. XLA cells (Btk deficient) PMA 0 ng/ml btk-mal-2 1 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait kinase_domain required to bind necessary binding region required to bind range range range range ph_domain required to bind necessary binding region required to bind range range range range n_terminus 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 physical association physical association Fig. 7B, 7C, 7D. Healthy/control cells btk-mal-3 2 8 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 6C-2 Healthy cells 100 (15 mins) mal-pik3r1-1 2 10 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 6E-2 XLA cells 100 (15 mins) mal-ptk2-1 3 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association Fig. 8C There is an interaction at the rafts of healthy/control neutrophils only after stimulation with PMA. Lyn, c-Src and Syk were associated with Mal in the cytosol of neutrophils from healthy controls but not in Btk-deficient neutrophils. XLA cells (Btk deficient) PMA 0 ng/ml mal-lyn 3 9 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association Fig. 8C-2 There is an interaction at the rafts of healthy/control neutrophils only after stimulation with PMA. Lyn, c-Src and Syk were associated with Mal in the cytosol of neutrophils from healthy controls but not in Btk-deficient neutrophils. Healthy control cells PMA 100 ng/ml for 5 mins mal-lyn-1 3 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey association association Fig. 8C-3 There is an interaction at the lipid rafts of healthy/control neutrophils only after stimulation with PMA. Lyn, c-Src and Syk were associated with Mal in the cytosol of neutrophils from healthy controls but not in Btk-deficient neutrophils. Healthy control cells PMA 0 OR 100 ng/ml for 5 mins mal-src-1 2 9 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 8D-2 XLA cells c-Src is associated with Mal in the cytoplasm but not in the membrane of normal resting neutrophils. (-in XLA cells Mal and c-Src localize at the membrane) 0