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/ mehraein_ghomi-2014-2 Targeting androgen receptor and JunD interaction for prevention of prostate cancer progression. in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps tag fluorescence tag visualisation by fluorescence tag fluorescence Mehraein-Ghomi F., Kegel S.J., Church D.R., Schmidt J.S., Reuter Q.R., Saphner E.L., Basu H.S., Wilding G. Cancer - Interactions investigated in the context of cancer Fluorescent polarization. AR-LBD(His-GST)(Rat Androgen-Receptor Ligand-Binding Domain)is added to a fluorescently-tagged androgen ligand (Fluormone) in the presence of competitor test compounds in microwell plates. The presence of effective competitors prevents the formation of a Fluormone/ AR-LBD(His-GST) complex resulting in a decrease of the polarization value due to ligand displacement caused by a competitor. Origins and details of constructs used obtained from Invitrogen website. gxw@medicine.wisc.edu Prostate (0270-4137) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. mehraein_ghomi-2014-1 Targeting androgen receptor and JunD interaction for prevention of prostate cancer progression. human-hep_3b Homo sapiens hepatocellular carcinoma (HCC) cells (p53-null). hep_3b human hepatocellular carcinoma (HCC) cells (p53-null). split luciferase complementation split luciferase complementation tag luciferase tag visualisation by luciferase assay tag luciferase Mehraein-Ghomi F., Kegel S.J., Church D.R., Schmidt J.S., Reuter Q.R., Saphner E.L., Basu H.S., Wilding G. Cancer - Interactions investigated in the context of cancer Gaussia luciferase reconstitution assay Gaussia luciferase is a naturally secreted protein by the copepod, Gaussia princeps. At 19kD, it is the smallest known luciferase and its catalytic properties make it one of the “brightest” known. (pmid:17099704) gxw@medicine.wisc.edu Prostate (0270-4137) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. mehraein_ghomi-2014-3 Targeting androgen receptor and JunD interaction for prevention of prostate cancer progression. human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Mehraein-Ghomi F., Kegel S.J., Church D.R., Schmidt J.S., Reuter Q.R., Saphner E.L., Basu H.S., Wilding G. Cancer - Interactions investigated in the context of cancer gxw@medicine.wisc.edu Prostate (0270-4137) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. bicalutamide Bicalutamide, an androgen antagonist small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) andr_rat Androgen receptor Ar Nr3c4 Dihydrotestosterone receptor Nuclear receptor subfamily 3 group C member 4 protein protein rat Rattus norvegicus (Rat) Rat MEVQLGLGRVYPRPPSKTYRGAFQNLFQSVREAIQNPGPRHPEAASIAPPGACLQQRQETSPRRRRRQQHPEDGSPQAHIRGTTGYLALEEEQQPSQQQSASEGHPESGCLPEPGAATAPGKGLPQQPPAPPDQDDSAAPSTLSLLGPTFPGLSSCSADIKDILSEAGTMQLLQQQQQQQQQQQQQQQQQQQQQQEVISEGSSSVRAREATGAPSSSKDSYLGGNSTISDSAKELCKAVSVSMGLGVEALEHLSPGEQLRGDCMYASLLGGPPAVRPTPCAPLAECKGLSLDEGPGKGTEETAEYSSFKGGYAKGLEGESLGCSGSSEAGSSGTLEIPSSLSLYKSGAVDEAAAYQNRDYYNFPLALSGPPHPPPPTHPHARIKLENPSDYGSAWAAAAAQCRYGDLASLHGGSVAGPSTGSPPATASSSWHTLFTAEEGQLYGPGGGGGSSSPSDAGPVAPYGYTRPPQGLASQEGDFSASEVWYPGGVVNRVPYPSPSCVKSEMGPWMENYSGPYGDMRLDSTRDHVLPIDYYFPPQKTCLICGDEASGCHYGALTCGSCKVFFKRAAEGKQKYLCASRNDCTIDKFRRKNCPSCRLRKCYEAGMTLGARKLKKLGNLKLQEEGENSSAGSPTEDPSQKMTVSHIEGYECQPIFLNVLEAIEPGVVCAGHDNNQPDSFAALLSSLNELGERQLVHVVKWAKALPGFRNLHVDDQMAVIQYSWMGLMVFAMGWRSFTNVNSRMLYFAPDLVFNEYRMHKSRMYSQCVRMRHLSQEFGWLQITPQEFLCMKALLLFSIIPVDGLKNQKFFDELRMNYIKELDRIIACKRKNPTSCSRRFYQLTKLLDSVQPIARELHQFTFDLLIKSHMVSVDFPEMMAEIISVQVPKILSGKVKPIYFHTQ fluormone_al_green Fluormone� AL Green, an androgen ligand, a fluorescent androgen analog. small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) a fluorescently-tagged androgen ligand (Fluormone� AL Green)- supplied by Invitrogen. It binds AR-LBD with a similar affinity as DHT. andr_human Androgen receptor Dihydrotestosterone receptor Nuclear receptor subfamily 3 group C member 4 AR DHTR NR3C4 protein protein human Homo sapiens Human MEVQLGLGRVYPRPPSKTYRGAFQNLFQSVREVIQNPGPRHPEAASAAPPGASLLLLQQQQQQQQQQQQQQQQQQQQQQQETSPRQQQQQQGEDGSPQAHRRGPTGYLVLDEEQQPSQPQSALECHPERGCVPEPGAAVAASKGLPQQLPAPPDEDDSAAPSTLSLLGPTFPGLSSCSADLKDILSEASTMQLLQQQQQEAVSEGSSSGRAREASGAPTSSKDNYLGGTSTISDNAKELCKAVSVSMGLGVEALEHLSPGEQLRGDCMYAPLLGVPPAVRPTPCAPLAECKGSLLDDSAGKSTEDTAEYSPFKGGYTKGLEGESLGCSGSAAAGSSGTLELPSTLSLYKSGALDEAAAYQSRDYYNFPLALAGPPPPPPPPHPHARIKLENPLDYGSAWAAAAAQCRYGDLASLHGAGAAGPGSGSPSAAASSSWHTLFTAEEGQLYGPCGGGGGGGGGGGGGGGGGGGGGGGEAGAVAPYGYTRPPQGLAGQESDFTAPDVWYPGGMVSRVPYPSPTCVKSEMGPWMDSYSGPYGDMRLETARDHVLPIDYYFPPQKTCLICGDEASGCHYGALTCGSCKVFFKRAAEGKQKYLCASRNDCTIDKFRRKNCPSCRLRKCYEAGMTLGARKLKKLGNLKLQEEGEASSTTSPTEETTQKLTVSHIEGYECQPIFLNVLEAIEPGVVCAGHDNNQPDSFAALLSSLNELGERQLVHVVKWAKALPGFRNLHVDDQMAVIQYSWMGLMVFAMGWRSFTNVNSRMLYFAPDLVFNEYRMHKSRMYSQCVRMRHLSQEFGWLQITPQEFLCMKALLLFSIIPVDGLKNQKFFDELRMNYIKELDRIIACKRKNPTSCSRRFYQLTKLLDSVQPIARELHQFTFDLLIKSHMVSVDFPEMMAEIISVQVPKILSGKVKPIYFHTQ jund_human Transcription factor jun-D JUND protein protein human Homo sapiens Human METPFYGDEALSGLGGGASGSGGSFASPGRLFPGAPPTAAAGSMMKKDALTLSLSEQVAAALKPAAAPPPTPLRADGAPSAAPPDGLLASPDLGLLKLASPELERLIIQSNGLVTTTPTSSQFLYPKVAASEEQEFAEGFVKALEDLHKQNQLGAGAAAAAAAAAAGGPSGTATGSAPPGELAPAAAAPEAPVYANLSSYAGGAGGAGGAATVAFAAEPVPFPPPPPPGALGPPRLAALKDEPQTVPDVPSFGESPPLSPIDMDTQERIKAERKRLRNRIAASKCRKRKLERISRLEEKVKTLKSQNTELASTASLLREQVAQLKQKVLSHVNSGCQLLPQHQVPAY ar-bicalutamide 1764903 n/a 1764906 competitor competitor competitor prey prey n/a 1764907 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false lbd domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 n/a 1764908 unspecified role unspecified role prey prey physical association physical association false false false Fig 2 Bicalutamide, an anti-androgen, is a "positive" control. The compound GWARJD22 also gave a decrease in polarisation values. ar-bicalutamide-1 1764903 n/a 1764907 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 lbd domain sufficient to bind sufficient binding region sufficient to bind range range range range false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1764908 unspecified role unspecified role prey prey physical association physical association false false false Fig 2 Bicalutamide, an anti-androgen, is a positive control. ar-jund 1764904 n/a 1764909 unspecified role unspecified role neutral component neutral component n_terminus_gluc1 (n-terminal part of of gaussia luciferase) luciferase tag luciferase tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1764910 unspecified role unspecified role neutral component neutral component c_terminus_gluc2 (c-terminal part of gaussia luciferase) luciferase tag luciferase tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 1 ar-jund-1 1764905 n/a 1764909 unspecified role unspecified role neutral component neutral component n/a 1764910 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false Fig. 4 synthetic androgen R1881 lead compound GWARJD10 (structure shown in fig 3)