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/ PAPA-1 Is a nuclear binding partner of IGFBP-2 and modulates its growth-promoting actions. PAPA-1 Is a nuclear binding partner of IGFBP-2 and modulates its growth-promoting actions. Mol. Endocrinol. (0888-8809) 2009 imex curation Miyako K., Cobb LJ., Francis M., Huang A., Peng B., Pintar JE., Ariga H., Cohen P. Cancer - Interactions investigated in the context of cancer hassy@mednet.ucla.edu 23-Feb-2010: Contacted by JYOTI. Human prostate Matchmaker cDNA library Only protein-protein interactions in vitro In vitro pull down pull down antibody detection identification by antibody antibody detection Miyako K., Cobb LJ., Francis M., Huang A., Peng B., Pintar JE., Ariga H., Cohen P. Cancer - Interactions investigated in the context of cancer Recombinant human IGFBP-2 used in this expt. was obtained from GroPep, vendor does not identify species for the expression system, given as 'mammalian'. hassy@mednet.ucla.edu Accepted 2009-Not a month!-4 by ORCHARD. 23-Feb-2010: Contacted by JYOTI. Human prostate Matchmaker cDNA library Mol. Endocrinol. (0888-8809) 2009 Only protein-protein interactions imex curation PAPA-1 Is a nuclear binding partner of IGFBP-2 and modulates its growth-promoting actions. PAPA-1 Is a nuclear binding partner of IGFBP-2 and modulates its growth-promoting actions. Mol. Endocrinol. (0888-8809) 2009 imex curation Miyako K., Cobb LJ., Francis M., Huang A., Peng B., Pintar JE., Ariga H., Cohen P. Cancer - Interactions investigated in the context of cancer hassy@mednet.ucla.edu 23-Feb-2010: Contacted by JYOTI. Human prostate Matchmaker cDNA library Only protein-protein interactions human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Miyako K., Cobb LJ., Francis M., Huang A., Peng B., Pintar JE., Ariga H., Cohen P. Cancer - Interactions investigated in the context of cancer hassy@mednet.ucla.edu Accepted 2009-Not a month!-4 by ORCHARD. 23-Feb-2010: Contacted by JYOTI. Human prostate Matchmaker cDNA library Mol. Endocrinol. (0888-8809) 2009 Only protein-protein interactions imex curation PAPA-1 Is a nuclear binding partner of IGFBP-2 and modulates its growth-promoting actions. PAPA-1 Is a nuclear binding partner of IGFBP-2 and modulates its growth-promoting actions. Mol. Endocrinol. (0888-8809) 2009 imex curation Miyako K., Cobb LJ., Francis M., Huang A., Peng B., Pintar JE., Ariga H., Cohen P. Cancer - Interactions investigated in the context of cancer hassy@mednet.ucla.edu 23-Feb-2010: Contacted by JYOTI. Human prostate Matchmaker cDNA library Only protein-protein interactions human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma confocal microscopy confocal microscopy immunostaining immunostaining Miyako K., Cobb LJ., Francis M., Huang A., Peng B., Pintar JE., Ariga H., Cohen P. Cancer - Interactions investigated in the context of cancer hassy@mednet.ucla.edu Accepted 2009-Not a month!-4 by ORCHARD. 23-Feb-2010: Contacted by JYOTI. Human prostate Matchmaker cDNA library Mol. Endocrinol. (0888-8809) 2009 orchard Only protein-protein interactions imex curation PAPA-1 Is a nuclear binding partner of IGFBP-2 and modulates its growth-promoting actions. PAPA-1 Is a nuclear binding partner of IGFBP-2 and modulates its growth-promoting actions. Mol. Endocrinol. (0888-8809) 2009 imex curation Miyako K., Cobb LJ., Francis M., Huang A., Peng B., Pintar JE., Ariga H., Cohen P. Cancer - Interactions investigated in the context of cancer hassy@mednet.ucla.edu 23-Feb-2010: Contacted by JYOTI. Human prostate Matchmaker cDNA library Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Miyako K., Cobb LJ., Francis M., Huang A., Peng B., Pintar JE., Ariga H., Cohen P. Cancer - Interactions investigated in the context of cancer hassy@mednet.ucla.edu Accepted 2009-Not a month!-4 by ORCHARD. 23-Feb-2010: Contacted by JYOTI. Human prostate Matchmaker cDNA library Mol. Endocrinol. (0888-8809) 2009 Only protein-protein interactions imex curation ibp2_human Insulin-like growth factor-binding protein 2 BP2 IBP2 IGFBP2 protein protein human Homo sapiens Human MLPRVGCPALPLPPPPLLPLLLLLLGASGGGGGARAEVLFRCPPCTPERLAACGPPPVAPPAAVAAVAGGARMPCAELVREPGCGCCSVCARLEGEACGVYTPRCGQGLRCYPHPGSELPLQALVMGEGTCEKRRDAEYGASPEQVADNGDDHSEGGLVENHVDSTMNMLGGGGSAGRKPLKSGMKELAVFREKVTEQHRQMGKGGKHHLGLEEPKKLRPPPARTPCQQELDQVLERISTMRLPDERGPLEHLYSLHIPNCDKHGLYNLKQCKMSLNGQRGECWCVNPNTGKLIQGAPTIRGDPECHLFYNEQQEARGVHTQRMQ 9E1436DBCCC6EA2A in80b_human INO80 complex subunit B PAP-1-associated protein 1 High mobility group AT-hook 1-like 4 INO80B HMGA1L4 PAPA1 ZNHIT4 Zinc finger HIT domain-containing protein 4 IES2 homolog protein protein human Homo sapiens Human MSKLWRRGSTSGAMEAPEPGEALELSLAGAHGHGVHKKKHKKHKKKHKKKHHQEEDAGPTQPSPAKPQLKLKIKLGGQVLGTKSVPTFTVIPEGPRSPSPLMVVDNEEEPMEGVPLEQYRAWLDEDSNLSPSPLRDLSGGLGGQEEEEEQRWLDALEKGELDDNGDLKKEINERLLTARQRALLQKARSQPSPMLPLPVAEGCPPPALTEEMLLKREERARKRRLQAARRAEEHKNQTIERLTKTAATSGRGGRGGARGERRGGRAAAPAPMVRYCSGAQGSTLSFPPGVPAPTAVSQRPSPSGPPPRCSVPGCPHPRRYACSRTGQALCSLQCYRINLQMRLGGPEGPGSPLLAT 235C836556B5F91F ibp2-in80b 4 5 unspecified role unspecified role bait bait mature peptide sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain 11 9 physical association physical association The IGFBP2 construct is described as the mature peptide so the signal sequence, as described in the UniProtKB entry, has been removed. ibp2-in80b-2 1 5 unspecified role unspecified role prey prey human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma 6 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1. Confirmation of IGFBP-2 and PAPA-1 interaction by GST pull down. ibp2-in80b-3 2 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig.2A Coimmunoprecipitation of IGFBP-2 and PAPA-1 in LNCaP cells. ibp2-in80b-5 3 6 unspecified role unspecified role neutral component neutral component 5 unspecified role unspecified role neutral component neutral component human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma colocalization colocalization Fig. 3. Nuclear colocalization of IGFBP-2 and PAPA-1 in LNCaP cells.