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/ Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. 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 Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu Accepted 2007-FEB-2 by DAVET. 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Biochem. Biophys. Res. Commun. (0006-291X) 2003 Only protein-protein interactions imex curation Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. 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 predetermined predetermined participant predetermined Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu Accepted 2007-FEB-2 by DAVET. 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Biochem. Biophys. Res. Commun. (0006-291X) 2003 Only protein-protein interactions imex curation Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast beta galactosidase beta galactosidase complementation beta galactosidase predetermined predetermined participant predetermined Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu Accepted 2007-FEB-2 by DAVET. 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Biochem. Biophys. Res. Commun. (0006-291X) 2003 Only protein-protein interactions imex curation Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Only protein-protein interactions human-c4_2 Homo sapiens c4-2 androgen-independent prostate cancer cells c4_2 Human androgen-independent prostate cancer cells C4-2 This cell line is also known as C4-2B. confocal microscopy confocal microscopy antibody detection identification by antibody antibody detection Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu Accepted 2007-FEB-2 by DAVET. 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Biochem. Biophys. Res. Commun. (0006-291X) 2003 Only protein-protein interactions imex curation Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Only protein-protein interactions human-c4_2 Homo sapiens c4-2 androgen-independent prostate cancer cells c4_2 Human androgen-independent prostate cancer cells C4-2 This cell line is also known as C4-2B. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu Accepted 2007-FEB-2 by DAVET. 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Biochem. Biophys. Res. Commun. (0006-291X) 2003 Only protein-protein interactions imex curation Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Only protein-protein interactions human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma confocal microscopy confocal microscopy antibody detection identification by antibody antibody detection Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu Accepted 2007-FEB-2 by DAVET. 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Biochem. Biophys. Res. Commun. (0006-291X) 2003 Only protein-protein interactions imex curation Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Metallothionein 2A interacts with the kinase domain of PKCmu in prostate cancer. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. 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 Rao PS., Jaggi M., Smith DJ., Hemstreet GP., Balaji KC. Cancer - Interactions investigated in the context of cancer kcbalaji@unmc.edu Accepted 2007-FEB-2 by DAVET. 02-MAR-2006: Contacted by JYOTI. Returned mail. A prostate cancer cDNA library was used. Biochem. Biophys. Res. Commun. (0006-291X) 2003 Only protein-protein interactions imex curation kpcd1_human Serine/threonine-protein kinase D1 nPKC-D1 Protein kinase D Protein kinase C mu type nPKC-mu PRKD1 PKD PKD1 PRKCM protein protein human Homo sapiens Human MSAPPVLRPPSPLLPVAAAAAAAAAALVPGSGPGPAPFLAPVAAPVGGISFHLQIGLSREPVLLLQDSSGDYSLAHVREMACSIVDQKFPECGFYGMYDKILLFRHDPTSENILQLVKAASDIQEGDLIEVVLSASATFEDFQIRPHALFVHSYRAPAFCDHCGEMLWGLVRQGLKCEGCGLNYHKRCAFKIPNNCSGVRRRRLSNVSLTGVSTIRTSSAELSTSAPDEPLLQKSPSESFIGREKRSNSQSYIGRPIHLDKILMSKVKVPHTFVIHSYTRPTVCQYCKKLLKGLFRQGLQCKDCRFNCHKRCAPKVPNNCLGEVTINGDLLSPGAESDVVMEEGSDDNDSERNSGLMDDMEEAMVQDAEMAMAECQNDSGEMQDPDPDHEDANRTISPSTSNNIPLMRVVQSVKHTKRKSSTVMKEGWMVHYTSKDTLRKRHYWRLDSKCITLFQNDTGSRYYKEIPLSEILSLEPVKTSALIPNGANPHCFEITTANVVYYVGENVVNPSSPSPNNSVLTSGVGADVARMWEIAIQHALMPVIPKGSSVGTGTNLHRDISVSISVSNCQIQENVDISTVYQIFPDEVLGSGQFGIVYGGKHRKTGRDVAIKIIDKLRFPTKQESQLRNEVAILQNLHHPGVVNLECMFETPERVFVVMEKLHGDMLEMILSSEKGRLPEHITKFLITQILVALRHLHFKNIVHCDLKPENVLLASADPFPQVKLCDFGFARIIGEKSFRRSVVGTPAYLAPEVLRNKGYNRSLDMWSVGVIIYVSLSGTFPFNEDEDIHDQIQNAAFMYPPNPWKEISHEAIDLINNLLQVKMRKRYSVDKTLSHPWLQDYQTWLDLRELECKIGERYITHESDDLRWEKYAGEQGLQYPTHLINPSASHSDTPETEETEMKALGERVSIL DIP protein Q15139 original sequence version: 79 DIP protein Q15139 original sequence version: 105 0BC9414C335D2DBB mt2_human Metallothionein-2 Metallothionein-II Metallothionein-2A MT2A CES1 MT2 protein protein human Homo sapiens Human MDPNCSCAAGDSCTCAGSCKCKECKCTSCKKSCCSCCPVGCAKCAQGCICKGASDKCSCCA 7B0A7E1F99843078 prkd1-mt2a 1 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association we report the results of our YHS screening experiment, which identified metallothionein 2A (MT 2A) as an interactor with the kinase domain of PKCl. prkd1-mt2a-1 2 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region C1 domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain c-terminal kinase domain (KD) sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association MT 2A interacted strongly with KD, relatively weakly with C1 domain, and failed to interact with C2, PH or KM domain MT 2A did not interact with the kinase mutant (K612W), C2 domain (271-320), or PH domain (409-552). 1 B prkd1-mt2a-2 3 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region C1 domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain c-terminal kinase domain (KD) sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association The strength of the interaction between the KD and C1 domain with MT 2A was tested using the liquid b-galactosidase assay. MT 2A showed strong interaction with KD and a relatively weak interaction with the C1 domain 1 B The interaction of MT 2A is stronger with the KD of PKCmu than the C1 domain. mt2a-prkd1 7 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 1 C The PKCl-MT 2A interaction in PC cells was confirmed by full-length PKCl co-IP with MT 2A mt2a-prkd1-1 5 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 1 C The PKCl-MT 2A interaction in PC cells was confirmed by full-length PKCl co-IP with MT 2A mt2a-prkd1-2 6 9 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization 2 Both PKCl and MT 2A co-localized in the cytoplasm, particularly to the perinuclear areas as shown by yellow areas in the merged images mt2a-prkd1-3 4 9 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization 2 Both PKCl and MT 2A co-localized in the cytoplasm, particularly to the perinuclear areas as shown by yellow areas in the merged images