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/ Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 Burbulis IE., Winkel-Shirley B. winkel@vt.edu 22-Jan-2008: Contacted by JYOTI. orchard arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Burbulis IE., Winkel-Shirley B. winkel@vt.edu winkel@vt.edu. Accepted 2007-DEC-20 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 orchard Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 Burbulis IE., Winkel-Shirley B. winkel@vt.edu 22-Jan-2008: Contacted by JYOTI. orchard in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Burbulis IE., Winkel-Shirley B. Arabidopsis wild-type (ecotypes Columbia (Co) or Landsberg (La)) and mutant types (lacking chalcone synthase or chalcone isomerase) seedlings were grown for 3 days under continuous white light in the presence of sucrose, conditions under which flavonoid enzymes accumulate to high levels, prior to assay. winkel@vt.edu winkel@vt.edu. Accepted 2007-DEC-20 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 orchard Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 Burbulis IE., Winkel-Shirley B. winkel@vt.edu 22-Jan-2008: Contacted by JYOTI. orchard 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 Burbulis IE., Winkel-Shirley B. winkel@vt.edu winkel@vt.edu. Accepted 2007-DEC-20 by ORCHARD. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 orchard chsy_arath Chalcone synthase Naringenin-chalcone synthase Protein TRANSPARENT TESTA 4 CHS TT4 MAC12.11 MAC12.14 At5g13930 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MVMAGASSLDEIRQAQRADGPAGILAIGTANPENHVLQAEYPDYYFRITNSEHMTDLKEKFKRMCDKSTIRKRHMHLTEEFLKENPHMCAYMAPSLDTRQDIVVVEVPKLGKEAAVKAIKEWGQPKSKITHVVFCTTSGVDMPGADYQLTKLLGLRPSVKRLMMYQQGCFAGGTVLRIAKDLAENNRGARVLVVCSEITAVTFRGPSDTHLDSLVGQALFSDGAAALIVGSDPDTSVGEKPIFEMVSAAQTILPDSDGAIDGHLREVGLTFHLLKDVPGLISKNIVKSLDEAFKPLGISDWNSLFWIAHPGGPAILDQVEIKLGLKEEKMRATRHVLSEYGNMSSACVLFILDEMRRKSAKDGVATTGEGLEWGVLFGFGPGLTVETVVLHSVPL 3FCED2BCF2CCAB0E dfra_arath Dihydroflavonol 4-reductase Dihydrokaempferol 4-reductase Protein TRANSPARENT TESTA 3 DFRA DFR TT3 MJB21.18 At5g42800 Flavanone 4-reductase protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MVSQKETVCVTGASGFIGSWLVMRLLERGYFVRATVRDPGNLKKVQHLLDLPNAKTLLTLWKADLSEEGSYDDAINGCDGVFHVATPMDFESKDPENEVIKPTVNGMLGIMKACVKAKTVRRFVFTSSAGTVNVEEHQKNVYDENDWSDLEFIMSKKMTGWMYFVSKTLAEKAAWDFAEEKGLDFISIIPTLVVGPFITTSMPPSLITALSPITRNEAHYSIIRQGQYVHLDDLCNAHIFLYEQAAAKGRYICSSHDATILTISKFLRPKYPEYNVPSTFEGVDENLKSIEFSSKKLTDMGFNFKYSLEEMFIESIETCRQKGFLPVSLSYQSISEIKTKNENIDVKTGDGLTDGMKPCNKTETGITGERTDAPMLAQQMCA 51F81F96512ECC3F cfi1_arath Chalcone--flavanone isomerase 1 Protein TRANSPARENT TESTA 5 CHI1 CFI TT5 T15C9_120 At3g55120 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSSNACASPSPFPAVTKLHVDSVTFVPSVKSPASSNPLFLGGAGVRGLDIQGKFVIFTVIGVYLEGNAVPSLSVKWKGKTTEELTESIPFFREIVTGAFEKFIKVTMKLPLTGQQYSEKVTENCVAIWKQLGLYTDCEAKAVEKFLEIFKEETFPPGSSILFALSPTGSLTVAFSKDDSIPETGIAVIENKLLAEAVLESIIGKNGVSPGTRLSVAERLSQLMMKNKDEKEVSDHSVEEKLAKEN 2EB68BF68D576B98 fl3h_arath Naringenin,2-oxoglutarate 3-dioxygenase Flavanone 3-hydroxylase F3H Protein TRANSPARENT TESTA 6 F3H TT6 F24M12.280 At3g51240 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAPGTLTELAGESKLNSKFVRDEDERPKVAYNVFSDEIPVISLAGIDDVDGKRGEICRQIVEACENWGIFQVVDHGVDTNLVADMTRLARDFFALPPEDKLRFDMSGGKKGGFIVSSHLQGEAVQDWREIVTYFSYPVRNRDYSRWPDKPEGWVKVTEEYSERLMSLACKLLEVLSEAMGLEKESLTNACVDMDQKIVVNYYPKCPQPDLTLGLKRHTDPGTITLLLQDQVGGLQATRDNGKTWITVQPVEGAFVVNLGDHGHFLSNGRFKNADHQAVVNSNSSRLSIATFQNPAPDATVYPLKVREGEKAILEEPITFAEMYKRKMGRDLELARLKKLAKEERDHKEVDKPVDQIFA 16BB0A3B318E5524 chi-chs 3 6 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey physical association physical association 2 no suitable source text dfr-chi 3 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association 2 no suitable source text chs-dfr 3 4 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association 2 no suitable source text chs-chi-f3h 2 7 unspecified role unspecified role prey prey arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress 4 unspecified role unspecified role bait bait region TRX tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress association association 3 Binding of CHI and F3H to TRX-CHS was also near the limit of detection. The authors noted that "binding of CHI and F3H to TRX-CHS was also near the limit of detection. However, using lysates from tt4 and tt5 seedlings, which lack CHS and CHI protein, respectively, significant amounts of CHS, CHI, and F3H could be eluted from the affinity columns. This suggests that the endogenous enzymes compete for binding with the recombinant protein immobilized on the affinity column". chi-chs-f3h 1 4 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association 4 This provides additional evidence for the association of CHS and F3H with CHI in the plant cell. chi-f3h-chs 2 7 unspecified role unspecified role prey prey arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress 4 unspecified role unspecified role prey prey region TRX tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress association association 3 A very small amount of CHS and F3H protein could be eluted reproducibly from the immobilized TRX-CHI by using lysates from wild-type seedlings A very small amount of CHS and F3H protein could be eluted reproducibly from the immobilized TRX-CHI by using lysates from wild-type seedlings