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/ p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). FASEB J. (0892-6638) 2013 imex curation Krishnaiah S.Y., Dodia C., Feinstein S.I., Fisher A.B. Affinomics - Interactions curated for the Affinomics consortium. abf@mail.med.upenn.edu 18-Sep-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2014-AUG-05 by ORCHARD FASEB J. (0892-6638) 2013 Only protein-protein interactions imex curation p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). FASEB J. (0892-6638) 2013 imex curation Krishnaiah S.Y., Dodia C., Feinstein S.I., Fisher A.B. Affinomics - Interactions curated for the Affinomics consortium. abf@mail.med.upenn.edu 18-Sep-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2014-AUG-05 by ORCHARD FASEB J. (0892-6638) 2013 Only protein-protein interactions imex curation p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). FASEB J. (0892-6638) 2013 imex curation Krishnaiah S.Y., Dodia C., Feinstein S.I., Fisher A.B. Affinomics - Interactions curated for the Affinomics consortium. abf@mail.med.upenn.edu 18-Sep-2014: Contacted by IntAct-Help. Only protein-protein interactions 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. please check I have calc-ed the Kd from the Ka correctly - inverse of it. abf@mail.med.upenn.edu Accepted 2014-AUG-05 by ORCHARD FASEB J. (0892-6638) 2013 Only protein-protein interactions imex curation p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). p67(phox) terminates the phospholipase A(2)-derived signal for activation of NADPH oxidase (NOX2). FASEB J. (0892-6638) 2013 imex curation Krishnaiah S.Y., Dodia C., Feinstein S.I., Fisher A.B. Affinomics - Interactions curated for the Affinomics consortium. abf@mail.med.upenn.edu 18-Sep-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2014-AUG-05 by ORCHARD FASEB J. (0892-6638) 2013 Only protein-protein interactions imex curation 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 017D955F0FEEDFBC 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 AECDEDD332858B8F 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 KILNVBDSWZSGLL-KXQOOQHDSA-O 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 663DB6B52D790F76 p67phox-prdx6 2 6 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 8 unspecified role unspecified role bait bait physical association physical association 1B Angiotensin 2 prdx6-p67phox 2 6 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 8 unspecified role unspecified role prey prey physical association physical association 1C Angiotensin 2 p67phox-prdx6-2 1 6 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 8 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction Figure 2 Angiotensin 2 U0126 - a MAP kinase inhibitor p67phox-prdx6-3 3 5 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 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 ecoli Escherichia coli (strain K12) 8 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 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 3 p67phox primarily binds to phosphoPrdx6, and this interaction is abolished by p67phox phosphorylation. 1_2_di_o_palmitoyl_sn_glycero_3_phosphocholine-p67phox 4 5 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 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 ecoli Escherichia coli (strain K12) 8 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 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 ecoli Escherichia coli (strain K12) 7 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 chemical synthesis Chemical synthesis (Chemical synthesis) phospholipase reaction phospholipase reaction 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 4 5 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 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 ecoli Escherichia coli (strain K12) 7 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 chemical synthesis Chemical synthesis (Chemical synthesis) phospholipase reaction phospholipase reaction 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.