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/ kolluri-2008-2 A short Nur77-derived peptide converts Bcl-2 from a protector to a killer. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Kolluri SK., Zhu X., Zhou X., Lin B., Chen Y., Sun K., Tian X., Town J., Cao X., Lin F., Zhai D., Kitada S., Luciano F., O'Donnell E., Cao Y., He F., Lin J., Reed JC., Satterthwait AC., Zhang XK. Apoptosis - Interactions involving proteins with a function related to apoptosis Bismaleimidohexane (BMH) cross-linker. Additional information on source of interacting proteins supplied by the authors asat@burnham.org, xzhang@burnham.org Cancer Cell (1535-6108) 2008 Only protein-protein interactions 16 imex curation Cancer - Interactions investigated in the context of cancer kolluri-2008-3 A short Nur77-derived peptide converts Bcl-2 from a protector to a killer. in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps predetermined predetermined participant predetermined Kolluri SK., Zhu X., Zhou X., Lin B., Chen Y., Sun K., Tian X., Town J., Cao X., Lin F., Zhai D., Kitada S., Luciano F., O'Donnell E., Cao Y., He F., Lin J., Reed JC., Satterthwait AC., Zhang XK. Apoptosis - Interactions involving proteins with a function related to apoptosis Bismaleimidohexane (BMH) cross-linker. Additional information on source of interacting proteins supplied by the authors asat@burnham.org, xzhang@burnham.org Cancer Cell (1535-6108) 2008 Only protein-protein interactions 16 imex curation Cancer - Interactions investigated in the context of cancer kolluri-2008-1 A short Nur77-derived peptide converts Bcl-2 from a protector to a killer. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Kolluri SK., Zhu X., Zhou X., Lin B., Chen Y., Sun K., Tian X., Town J., Cao X., Lin F., Zhai D., Kitada S., Luciano F., O'Donnell E., Cao Y., He F., Lin J., Reed JC., Satterthwait AC., Zhang XK. Apoptosis - Interactions involving proteins with a function related to apoptosis Bismaleimidohexane (BMH) cross-linker. Additional information on source of interacting proteins supplied by the authors asat@burnham.org, xzhang@burnham.org Cancer Cell (1535-6108) 2008 Only protein-protein interactions 16 imex curation Cancer - Interactions investigated in the context of cancer kolluri-2008-4 A short Nur77-derived peptide converts Bcl-2 from a protector to a killer. in vitro In vitro cd circular dichroism CD CD cd predetermined predetermined participant predetermined Kolluri SK., Zhu X., Zhou X., Lin B., Chen Y., Sun K., Tian X., Town J., Cao X., Lin F., Zhai D., Kitada S., Luciano F., O'Donnell E., Cao Y., He F., Lin J., Reed JC., Satterthwait AC., Zhang XK. Apoptosis - Interactions involving proteins with a function related to apoptosis Bismaleimidohexane (BMH) cross-linker. Additional information on source of interacting proteins supplied by the authors asat@burnham.org, xzhang@burnham.org Cancer Cell (1535-6108) 2008 Only protein-protein interactions 16 imex curation Cancer - Interactions investigated in the context of cancer kolluri-2008-5 A short Nur77-derived peptide converts Bcl-2 from a protector to a killer. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Kolluri SK., Zhu X., Zhou X., Lin B., Chen Y., Sun K., Tian X., Town J., Cao X., Lin F., Zhai D., Kitada S., Luciano F., O'Donnell E., Cao Y., He F., Lin J., Reed JC., Satterthwait AC., Zhang XK. Apoptosis - Interactions involving proteins with a function related to apoptosis Bismaleimidohexane (BMH) cross-linker. Additional information on source of interacting proteins supplied by the authors asat@burnham.org, xzhang@burnham.org Cancer Cell (1535-6108) 2008 Only protein-protein interactions 16 imex curation Cancer - Interactions investigated in the context of cancer kolluri-2008-6 A short Nur77-derived peptide converts Bcl-2 from a protector to a killer. human-dohh2 Human DOHH2 lymphoma follicular B cell dohh2 DoHH2 lymphoma, follicular, B cells crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Kolluri SK., Zhu X., Zhou X., Lin B., Chen Y., Sun K., Tian X., Town J., Cao X., Lin F., Zhai D., Kitada S., Luciano F., O'Donnell E., Cao Y., He F., Lin J., Reed JC., Satterthwait AC., Zhang XK. Apoptosis - Interactions involving proteins with a function related to apoptosis Bismaleimidohexane (BMH) cross-linker. Additional information on source of interacting proteins supplied by the authors asat@burnham.org, xzhang@burnham.org Cancer Cell (1535-6108) 2008 Only protein-protein interactions 16 imex curation Cancer - Interactions investigated in the context of cancer bcl2_human Apoptosis regulator Bcl-2 BCL2 protein protein human Homo sapiens Human MAHAGRTGYDNREIVMKYIHYKLSQRGYEWDAGDVGAAPPGAAPAPGIFSSQPGHTPHPAASRDPVARTSPLQTPAAPGAAAGPALSPVPPVVHLTLRQAGDDFSRRYRRDFAEMSSQLHLTPFTARGRFATVVEELFRDGVNWGRIVAFFEFGGVMCVESVNREMSPLVDNIALWMTEYLNRHLHTWIQDNGGWDAFVELYGPSMRPLFDFSWLSLKTLLSLALVGACITLGAYLGHK nr4a1_human Nuclear receptor subfamily 4 group A member 1 Orphan nuclear receptor HMR Early response protein NAK1 Orphan nuclear receptor TR3 ST-59 GFRP1 HMR NAK1 NR4A1 Nuclear hormone receptor NUR/77 Testicular receptor 3 protein protein human Homo sapiens Human MPCIQAQYGTPAPSPGPRDHLASDPLTPEFIKPTMDLASPEAAPAAPTALPSFSTFMDGYTGEFDTFLYQLPGTVQPCSSASSSASSTSSSSATSPASASFKFEDFQVYGCYPGPLSGPVDEALSSSGSDYYGSPCSAPSPSTPSFQPPQLSPWDGSFGHFSPSQTYEGLRAWTEQLPKASGPPQPPAFFSFSPPTGPSPSLAQSPLKLFPSQATHQLGEGESYSMPTAFPGLAPTSPHLEGSGILDTPVTSTKARSGAPGGSEGRCAVCGDNASCQHYGVRTCEGCKGFFKRTVQKNAKYICLANKDCPVDKRRRNRCQFCRFQKCLAVGMVKEVVRTDSLKGRRGRLPSKPKQPPDASPANLLTSLVRAHLDSGPSTAKLDYSKFQELVLPHFGKEDAGDVQQFYDLLSGSLEVIRKWAEKIPGFAELSPADQDLLLESAFLELFILRLAYRSKPGEGKLIFCSGLVLHRLQCARGFGDWIDSILAFSRSLHSLLVDVPAFACLSALVLITDRHGLQEPRRVEELQNRIASCLKEHVAAVAGEPQPASCLSRLLGKLPELRTLCTQGLQRIFYLKLEDLVPPPPIIDKIFMDTLPF b2l10_human Bcl-2-like protein 10 BCL2L10 BCLB Apoptosis regulator Bcl-B Anti-apoptotic protein NrH protein protein human Homo sapiens Human MVDQLRERTTMADPLRERTELLLADYLGYCAREPGTPEPAPSTPEAAVLRSAAARLRQIHRSFFSAYLGYPGNRFELVALMADSVLSDSPGPTWGRVVTLVTFAGTLLERGPLVTARWKKWGFQPRLKEQEGDVARDCQRLVALLSSRLMGQHRAWLQAQGGWDGFCHFFRTPFPLAFWRKQLVQAFLSCLLTTAFIYLWTRLL b2la1_human Bcl-2-related protein A1 Protein BFL-1 Hemopoietic-specific early response protein Protein GRS BCL2A1 BCL2L5 BFL1 GRS HBPA1 Bcl-2-like protein 5 protein protein human Homo sapiens Human MTDCEFGYIYRLAQDYLQCVLQIPQPGSGPSKTSRVLQNVAFSVQKEVEKNLKSCLDNVNVVSVDTARTLFNQVMEKEFEDGIINWGRIVTIFAFEGILIKKLLRQQIAPDVDTYKEISYFVAEFIMNNTGEWIRQNGGWENGFVKKFEPKSGWMTFLEVTGKICEMLSLLKQYC q07817-1 Bcl-2-like protein 1 BCLX Apoptosis regulator Bcl-X BCL2L1 BCL2L Bcl-xL protein protein human Homo sapiens Human MSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEMETPSAINGNPSWHLADSPAVNGATGHSSSLDAREVIPMAAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVNWGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQERFNRWFLTGMTVAGVVLLGSLFSRK bax_human Apoptosis regulator BAX BAX BCL2L4 Bcl-2-like protein 4 protein protein human Homo sapiens Human MDGSGEQPRGGGPTSSEQIMKTGALLLQGFIQDRAGRMGGEAPELALDPVPQDASTKKLSECLKRIGDELDSNMELQRMIAAVDTDSPREVFFRVAADMFSDGNFNWGRVVALFYFASKLVLKALCTKVPELIRTIMGWTLDFLRERLLGWIQDQGGWDGLLSYFGTPTWQTVTIFVAGVLTASLTIWKKMG nr4a1-bcl2-3 Interaction between NR4A1 and BCL2 demonstrated by coimmunopreciptiation 782490 n/a 782496 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P10415:p.Thr69_Ser70delinsGluGlu mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false undefined mutation mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing insertion analysis insertion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 782497 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 782510 782505 physical association physical association false false false Figs. 3C, 5A, 5G P10415; P22736 transfection Exact insertion point of the insertion mutant into the BCL-2 interacting region is not given in the text. nr4a1-bcl2l10-1 Interaction between NR4A1 and BCL2L10 demonstrated by coimmunopreciptiation 782490 n/a 782497 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 782498 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 3C Q9HD36; P22736 transfection nr4a1-bcl2la1-1 Interaction between NR4A1 and BCL2LA1 demonstrated by coimmunopreciptiation 782490 n/a 782499 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 782497 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 3C Q16548; P22736 transfection bcl2-1 Homodimerization of BCL2 782490 n/a 782496 unspecified role unspecified role bait bait region delta bh4 sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range false range range certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 782496 unspecified role unspecified role prey prey bh4 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 7A The BH4 domain could not bind full-length Bcl-2, likely due to inaccessibility of a BH4-binding site in the C-terminal region Coordinates of the deletion mutants not given by the author so have been inferred from the UniProtKB entry. bcl2-2 Homodimerization of BCL2 782490 n/a 782496 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 782496 unspecified role unspecified role prey prey bh4 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figs. 7B, 7C NR4A1 - 489-497. Interaction of BH4 domain with full length protein only occurs in the presence of this agonist, suggesting that binding of NR4A1 with Bcl-2 reorganizes Bcl-2, leading to the exposure of the BH4-binding site in the C-terminal region. nr4a1-bcl2-4 Interaction between NR4A1 and BCL2 demonstrated by fluorescence polarisation 782491 n/a 782496 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 782497 unspecified role unspecified role prey prey region fitc labelled fluorescein isothiocyanate label fluorescein isothiocyanate labbeled FITC labelled FITC labelled fluorescein isothiocyanate labbeled fitc labelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P22736:p.[Phe489Ala;Leu497Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false Figs. 3D, 4E, 5B, 5H Replacing L-amino acids of P22736 with D-amino acids did not diminish its effect. pH=7.6, Temp=25C nr4a1-bcl2-5 Interaction between NR4A1 and BCL2 demonstrated by fluorescence polarisation 782491 n/a 782497 competitor competitor competitor prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P22736:p.[Phe489Ala;Leu497Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 782496 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 782497 unspecified role unspecified role prey prey region fitc labelled fluorescein isothiocyanate label fluorescein isothiocyanate labbeled FITC labelled FITC labelled fluorescein isothiocyanate labbeled fitc labelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) association association false false false Figs. 3E, 4F, 5B, 5H Replacing L-amino acids of P22736 with D-amino acids did not diminish the effect of P22736. Kd of D-enantiomer of P22736 = 1.51E-7M EC50 inhibition by D-enantiomer of P22736 = 4.5E-6M, Ki=1.9E-6M EC50 inhibition by P22736 = 3.2E-6M, Ki=1.2E-6M pH=7.6, Temp=20C bcl2-bclx-1 Interaction between BCL2 and BCLX demonstrated by fluorescence polarization 782491 n/a 782500 unspecified role unspecified role neutral component neutral component n/a 782496 unspecified role unspecified role neutral component neutral component bh3 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region F5M fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false pH = 7.4, Temp. = 25C Fig. 8D nr4a1-bcl2-1 Interaction between NR4A1 and BCL2 demonstrated by coimmunopreciptiation 782492 n/a 782497 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 782496 unspecified role unspecified role bait bait physical association physical association false false false Fig. 3A P10415; P22736 transfection nr4a1-bcl2-2 Interaction between NR4A1 and BCL2 demonstrated by coimmunopreciptiation 782492 n/a 782497 competitor competitor competitor prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region D-arg octomer tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 782496 unspecified role unspecified role bait bait n/a 782497 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false association association false false false Fig. 3B P10415; P22736 transfection Replacing L-amino acids with D-amino acids did not diminish the effect of P22736 . nr4a1-bcl2-6 Interaction between NR4A1 and BCL2 demonstrated by circular dichroism 782493 n/a 782496 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 n/a 782497 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P22736:p.[Phe489Ala;Leu497Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 direct interaction direct interaction false false false Fig. 4C, 4d, 4E, 5C, 5D, Supp. Fig. 8 Replacing L-amino acids of P22736 with D-amino acids did not diminish its effect. pH=7.6, Temp=25C Binding of D-enantiomer of P22736 Kd=2.5E-6M nr4a1-bcl2-7 Interaction between NR4A1 and BCL2 demonstrated by circular dichroism 782493 n/a 782496 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 n/a 782497 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 direct interaction direct interaction false false false Fig. 5I bax-1 Homodimerization of BAX 782494 n/a 782501 unspecified role unspecified role neutral component neutral component Stoichiometry: 2.0 physical association physical association false false false Homodimerization identified by a calibrated change in molecular weight of the protein. Fig. 6A, 6B NR4A1 (P22736), amino acids 489-497. Replacing the L-amino acids with D-amino acids did not diminish its effect. Stimulation by NR4A1 required the presence of BCL2. bax-2 Homotrimerization of BAX 782494 n/a 782501 unspecified role unspecified role neutral component neutral component Stoichiometry: 3.0 physical association physical association false false false Homotrimerization identified by a calibrated change in molecular weight of the protein. Fig. 6A, 6B NR4A1 (P22736), amino acids 489-497. Replacing the L-amino acids with D-amino acids did not diminish its effect. Stimulation by NR4A1 required the presence of BCL2. bax-3 Homotrimerization of BAX 782495 n/a 782501 unspecified role unspecified role neutral component neutral component Stoichiometry: 3.0 physical association physical association false false false Homotrimerization identified by a calibrated change in molecular weight of the protein. Fig. 6C NR4A1 (P22736), amino acids 489-497. Replacing the L-amino acids with D-amino acids did not diminish its effect. DoHH2 cells express high levels of BCL2. bax-1 Homodimerization of BAX 782495 n/a 782501 unspecified role unspecified role neutral component neutral component Stoichiometry: 2.0 physical association physical association false false false Homodimerization identified by a calibrated change in molecular weight of the protein. Fig. 6C NR4A1 (P22736), amino acids 489-497. Replacing the L-amino acids with D-amino acids did not diminish its effect. DoHH2 cells express high levels of BCL2.