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/ http://www.ebi.ac.uk/ krishnaiah-2013-1 p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). mouse-endothelium Mus musculus endothelium Endothelial cells [endothelium] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Krishnaiah S.Y., Dodia C., Feinstein S.I., Fisher A.B. Affinomics - Interactions curated for the Affinomics consortium. MPMVECs- mouse pulmonary microvascular endothelial cells. abf@mail.med.upenn.edu FASEB J. (0892-6638) 2013 Only protein-protein interactions imex curation krishnaiah-2013-4 p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). in vitro In vitro phospholipase assay phospholipase assay tag visualisation tag visualisation Krishnaiah S.Y., Dodia C., Feinstein S.I., Fisher A.B. Affinomics - Interactions curated for the Affinomics consortium. Mixed unilamellar liposomes containing DPPC (1,2-di-O-palmitoyl-sn-glycero-3-phosphocholine) with a [9,10-3H]-palmitate radio-label as a substrate. Liposomes consist of 3H-DPPC, egg phosphatidylcholine, phosphatidylglycerol, and cholesterol. After incubation of liposomes and proteins at 37degrees the radiolabeled free fatty acid product was extracted, resolved by thin-layer chromatography and analyzed by scintillation counting. abf@mail.med.upenn.edu FASEB J. (0892-6638) 2013 Only protein-protein interactions imex curation krishnaiah-2013-2 p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). mouse-endothelium Mus musculus endothelium Endothelial cells [endothelium] pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Krishnaiah S.Y., Dodia C., Feinstein S.I., Fisher A.B. Affinomics - Interactions curated for the Affinomics consortium. MPMVECs- mouse pulmonary microvascular endothelial cells. abf@mail.med.upenn.edu FASEB J. (0892-6638) 2013 Only protein-protein interactions imex curation krishnaiah-2013-3 p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Krishnaiah S.Y., Dodia C., Feinstein S.I., Fisher A.B. Affinomics - Interactions curated for the Affinomics consortium. abf@mail.med.upenn.edu FASEB J. (0892-6638) 2013 Only protein-protein interactions imex curation prdx6_mouse Peroxiredoxin-6 Lysophosphatidylcholine acyltransferase 5 Glutathione-dependent peroxiredoxin Antioxidant protein 2 1-Cys peroxiredoxin Acidic calcium-independent phospholipase A2 Non-selenium glutathione peroxidase Prdx6 Aop2 Ltw4 Prdx5 protein protein mouse Mus musculus Mouse MPGGLLLGDEAPNFEANTTIGRIRFHDFLGDSWGILFSHPRDFTPVCTTELGRAAKLAPEFAKRNVKLIALSIDSVEDHLAWSKDINAYNGETPTEKLPFPIIDDKGRDLAILLGMLDPVEKDDNNMPVTARVVFIFGPDKKLKLSILYPATTGRNFDEILRVVDSLQLTGTKPVATPVDWKKGESVMVVPTLSEEEAKQCFPKGVFTKELPSGKKYLRYTPQP ncf2_mouse Neutrophil cytosol factor 2 Ncf2 Noxa2 P67phox p67-phox 67 kDa neutrophil oxidase factor NADPH oxidase activator 2 Neutrophil NADPH oxidase factor 2 protein protein mouse Mus musculus Mouse MSLAEAIRLWNEGVLAADKKDWKGALEAFSEVQDPHSRICFNIGCVNTILENLQAAEQAFTKSINRDKHSAVAYFQRGMLYYRMEKYDLAIKDLKEALTQLRGNQLIDYKILGLQFKLFACEVLYNIALMHAKKEEWKKAEEQLALATNMKSEPRHSKIDKAMESIWKQKLFEPVVIPVGRLFRPNERQVAQLAKKDYLGKATVVASVVHQDNFSGFAPLQPQSAEPPPRPKTPEIFRALEGEAHRVLFGFVPETPEELQVMPGNIVFVLKKGSDNWATVMFNGQKGLVPCNYLEPVELRIHPQSQPQEDTSPESDIPPPPNSSPPGRLQLSPGHKQKEPKELKLSVPMPYMLKVHYKYTVVMETRLGLPYSQLRNMVSKKLALSPEHTKLSYRRRDSHELLLLSEESMKDAWGQVKNYCLTLWCEHTVGDQGLIDEPIQRENSDASKQTTEPQPKEGTQVVAIFSYEAAQPEDLEFVEGDVILVLSHVNEEWLEGECKGKVGIFPKAFVEGCAAKNLEGIPREV prdx6_human Peroxiredoxin-6 PRDX6 AOP2 KIAA0106 Antioxidant protein 2 1-Cys peroxiredoxin Acidic calcium-independent phospholipase A2 Non-selenium glutathione peroxidase 24 kDa protein Liver 2D page spot 40 Red blood cells page spot 12 Glutathione-dependent peroxiredoxin Lysophosphatidylcholine acyltransferase 5 protein protein human Homo sapiens Human MPGGLLLGDVAPNFEANTTVGRIRFHDFLGDSWGILFSHPRDFTPVCTTELGRAAKLAPEFAKRNVKLIALSIDSVEDHLAWSKDINAYNCEEPTEKLPFPIIDDRNRELAILLGMLDPAEKDEKGMPVTARVVFVFGPDKKLKLSILYPATTGRNFDEILRVVISLQLTAEKRVATPVDWKDGDSVMVLPTIPEEEAKKLFPKGVFTKELPSGKKYLRYTPQP 1,2-di-o-palmitoyl-sn-glycero-3-phosphocholine 1,2-di-O-palmitoyl-sn-glycero-3-phosphocholine 1,2-di-O-hexadecanoyl-sn-glycero-3-phosphocholine 1,2-Dipalmitoyl-3-sn-phosphatidylcholine 1,2-Dipalmitoyl-L-lecithin 1,2-dipalmitoyl-sn-glycero-3-phosphocholine Dipalmitoyl phosphatidylcholine DPPC PC(16:0/16:0) (7R)-4-hydroxy-N,N,N-trimethyl-10-oxo-7-(hexadecanoyloxy)-3,5,9-trioxa-4-phosphapentacosan-1-aminium 4-oxide small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) KILNVBDSWZSGLL-KXQOOQHDSA-O p67phox-prdx6 1546637 n/a 1546641 unspecified role unspecified role prey prey required_phosphores phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1546642 unspecified role unspecified role bait bait physical association physical association false false false 1B Angiotensin 2 prdx6-p67phox 1546637 n/a 1546641 unspecified role unspecified role bait bait required_phosphores phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1546642 unspecified role unspecified role prey prey physical association physical association false false false 1C Angiotensin 2 1_2_di_o_palmitoyl_sn_glycero_3_phosphocholine-p67phox 1546638 n/a 1546643 enzyme enzyme neutral component neutral component c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false required_phosphores phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1546642 inhibitor inhibitor neutral component neutral component thr233 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain false c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1546644 enzyme target enzyme target substrate substrate neutral component neutral component 2-[9,10-3h]-palmitate 3h radiolabel 3h radiolabel 3H tritium H3 certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phospholipase reaction phospholipase reaction false false false Figure 4 The addition of p67phox inhibited the PLA2 activity of both native and phosphoPrdx6, while phosphorylated p67phox had no effect. p67-phox inhibits non-competitively. 1_2_di_o_palmitoyl_sn_glycero_3_phosphocholine-ncf2 1546638 n/a 1546643 enzyme enzyme neutral component neutral component required_phosphores phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1546644 enzyme target enzyme target substrate substrate neutral component neutral component 2-[9,10-3h]-palmitate 3h radiolabel 3h radiolabel 3H tritium H3 certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phospholipase reaction phospholipase reaction false false false Figure 4-2 The addition of p67phox inhibited the PLA2 activity of both native and phosphoPrdx6, while phosphorylated p67phox had no effect. p67-phox inhibits non-competitively. p67phox-prdx6-2 1546639 n/a 1546641 unspecified role unspecified role neutral component neutral component required_phosphores phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1546642 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 2 Angiotensin 2 U0126 - a MAP kinase inhibitor p67phox-prdx6-3 1546640 n/a 1546643 unspecified role unspecified role neutral component neutral component required_phosphores phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1546642 unspecified role unspecified role neutral component neutral component thr-233 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain false c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1546640 Figure 3 p67phox primarily binds to phosphoPrdx6, and this interaction is abolished by p67phox phosphorylation.