InnateDB InnateDB honda-2012-1 The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils. 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 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 honda-2012-2 The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils. 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 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 honda-2012-3 The kinase Btk negatively regulates the production of reactive oxygen species and stimulation-induced apoptosis in human neutrophils. 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 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 rac2_human Ras-related C3 botulinum toxin substrate 2 p21-Rac2 Small G protein GX RAC2 protein protein human Homo sapiens Human MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVMVDSKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASYENVRAKWFPEVRHHCPSTPIILVGTKLDLRDDKDTIEKLKEKKLAPITYPQGLALAKEIDSVKYLECSALTQRGLKTVFDEAIRAVLCPQPTRQQKRACSLL mal_human Myelin and lymphocyte protein MAL T-lymphocyte maturation-associated protein protein protein human Homo sapiens Human MAPAAATGGSTLPSGFSVFTTLPDLLFIFEFIFGGLVWILVASSLVPWPLVQGWVMFVSVFCFVATTTLIILYIIGAHGGETSWVTLDAAYHCTAALFYLSASVLEALATITMQDGFTYRHYHENIAAVVFSYIATLLYVVHAVFSLIRWKSS 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 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 src_human Proto-oncogene tyrosine-protein kinase Src pp60c-src SRC SRC1 Proto-oncogene c-Src protein protein human Homo sapiens Human MGSNKSKPKDASQRRRSLEPAENVHGAGGGAFPASQTPSKPASADGHRGPSAAFAPAAAEPKLFGGFNSSDTVTSPQRAGPLAGGVTTFVALYDYESRTETDLSFKKGERLQIVNNTEGDWWLAHSLSTGQTGYIPSNYVAPSDSIQAEEWYFGKITRRESERLLLNAENPRGTFLVRESETTKGAYCLSVSDFDNAKGLNVKHYKIRKLDSGGFYITSRTQFNSLQQLVAYYSKHADGLCHRLTTVCPTSKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTRVAIKTLKPGTMSPEAFLQEAQVMKKLRHEKLVQLYAVVSEEPIYIVTEYMSKGSLLDFLKGETGKYLRLPQLVDMAAQIASGMAYVERMNYVHRDLRAANILVGENLVCKVADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELTTKGRVPYPGMVNREVLDQVERGYRMPCPPECPESLHDLMCQCWRKEPEERPTFEYLQAFLEDYFTSTEPQYQPGENL 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 ksyk_human Tyrosine-protein kinase SYK SYK Spleen tyrosine kinase p72-Syk protein protein human Homo sapiens Human MASSGMADSANHLPFFFGNITREEAEDYLVQGGMSDGLYLLRQSRNYLGGFALSVAHGRKAHHYTIERELNGTYAIAGGRTHASPADLCHYHSQESDGLVCLLKKPFNRPQGVQPKTGPFEDLKENLIREYVKQTWNLQGQALEQAIISQKPQLEKLIATTAHEKMPWFHGKISREESEQIVLIGSKTNGKFLIRARDNNGSYALCLLHEGKVLHYRIDKDKTGKLSIPEGKKFDTLWQLVEHYSYKADGLLRVLTVPCQKIGTQGNVNFGGRPQLPGSHPATWSAGGIISRIKSYSFPKPGHRKSSPAQGNRQESTVSFNPYEPELAPWAADKGPQREALPMDTEVYESPYADPEEIRPKEVYLDRKLLTLEDKELGSGNFGTVKKGYYQMKKVVKTVAVKILKNEANDPALKDELLAEANVMQQLDNPYIVRMIGICEAESWMLVMEMAELGPLNKYLQQNRHVKDKNIIELVHQVSMGMKYLEESNFVHRDLAARNVLLVTQHYAKISDFGLSKALRADENYYKAQTHGKWPVKWYAPECINYYKFSSKSDVWSFGVLMWEAFSYGQKPYRGMKGSEVTAMLEKGERMGCPAGCPREMYDLMNLCWTYDVENRPGFAAVELRLRNYYYDVVN 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 cybb-rac2 1308778 n/a 1308781 unspecified role unspecified role neutral component neutral component n/a 1308782 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 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 1308778 n/a 1308783 unspecified role unspecified role neutral component neutral component n/a 1308784 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 0 Fig. 6C. Healthy cells btk-mal-3 1308778 n/a 1308784 unspecified role unspecified role neutral component neutral component n/a 1308783 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 6C-2 Healthy cells 100 (15 mins) mal-pik3r1 1308778 n/a 1308783 unspecified role unspecified role neutral component neutral component n/a 1308785 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 100 (15 mins) Fig. 6E. Healthy cells mal-pik3r1-1 1308778 n/a 1308785 unspecified role unspecified role neutral component neutral component n/a 1308783 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 6E-2 XLA cells 100 (15 mins) mal-src 1308778 n/a 1308786 unspecified role unspecified role neutral component neutral component n/a 1308783 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 8D. Healthy/control cells c-Src is associated with Mal in the cytoplasm but not in the membrane of normal resting neutrophils. 0 mal-src-1 1308778 n/a 1308783 unspecified role unspecified role neutral component neutral component n/a 1308786 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 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 btk-mal-1 1308779 n/a 1308784 unspecified role unspecified role bait bait n/a 1308783 unspecified role unspecified role prey prey physical association physical association false false false Fig. 6A. PMA 0 ng/ml Healthy cells This interaction was not observed in healthy cells after PMA stimulation. mal-btk 1308779 n/a 1308783 unspecified role unspecified role bait bait n/a 1308784 unspecified role unspecified role prey prey physical association physical association false false false Fig. 6B. Healthy control cells PMA - 0 ng/ml pik3r1-mal 1308779 n/a 1308785 unspecified role unspecified role bait bait n/a 1308783 unspecified role unspecified role prey prey physical association physical association false false false Fig. 6D. XLA (Btk deficient) neutrophils. Interaction detected only in XLA neutrophils. mal-ptk2 1308779 n/a 1308787 unspecified role unspecified role prey prey n/a 1308788 unspecified role unspecified role prey prey n/a 1308783 unspecified role unspecified role bait bait n/a 1308786 unspecified role unspecified role prey prey association association false false false Fig. S6. Interaction detected only in XLA neutrophils. XLA cells (Btk deficient) PMA 0 ng/ml mal-ptk2-1 1308779 n/a 1308789 unspecified role unspecified role prey prey n/a 1308788 unspecified role unspecified role prey prey n/a 1308783 unspecified role unspecified role bait bait n/a 1308786 unspecified role unspecified role prey prey association association false false false 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 1308779 n/a 1308783 unspecified role unspecified role bait bait n/a 1308789 unspecified role unspecified role prey prey n/a 1308786 unspecified role unspecified role prey prey n/a 1308788 unspecified role unspecified role prey prey association association false false false 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 1308779 n/a 1308789 unspecified role unspecified role prey prey n/a 1308786 unspecified role unspecified role prey prey n/a 1308783 unspecified role unspecified role bait bait n/a 1308788 unspecified role unspecified role prey prey association association false false false 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 btk-mal-2 1308780 n/a 1308783 unspecified role unspecified role prey prey n/a 1308784 unspecified role unspecified role bait bait kinase_domain required to bind necessary binding region required to bind range range range range false ph_domain required to bind necessary binding region required to bind range range range range false 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 false physical association physical association false false false Fig. 7B, 7C, 7D. Healthy/control cells