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/ A chloroplast FKBP interacts with and affects the accumulation of Rieske subunit of cytochrome bf complex. A chloroplast FKBP interacts with and affects the accumulation of Rieske subunit of cytochrome bf complex. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 imex curation Gupta R., Mould RM., He Z., Luan S. sluan@nature.berkeley.edu Arabidopsis ACT cDNA expression library CD4-22. Description: Generated from mRNA isolated from 3 day-old etiolated Columbia seedlings; library consists of 36,000,000 independent recombinants and was amplified on LE392; 22 of 23 clones tested contained inserts. orchard 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 Gupta R., Mould RM., He Z., Luan S. sluan@nature.berkeley.edu. sluan@nature.berkeley.edu Accepted 2009-NOV-9 by ORCHARD. Arabidopsis ACT cDNA expression library CD4-22. Description: Generated from mRNA isolated from 3 day-old etiolated Columbia seedlings; library consists of 36,000,000 independent recombinants and was amplified on LE392; 22 of 23 clones tested contained inserts. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Only protein-protein interactions imex curation A chloroplast FKBP interacts with and affects the accumulation of Rieske subunit of cytochrome bf complex. A chloroplast FKBP interacts with and affects the accumulation of Rieske subunit of cytochrome bf complex. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 imex curation Gupta R., Mould RM., He Z., Luan S. sluan@nature.berkeley.edu Arabidopsis ACT cDNA expression library CD4-22. Description: Generated from mRNA isolated from 3 day-old etiolated Columbia seedlings; library consists of 36,000,000 independent recombinants and was amplified on LE392; 22 of 23 clones tested contained inserts. orchard Only protein-protein interactions human-ws1 Homo sapiens ws1 Human fibroblast derived from the midscapular skin of a 12 week Negro female embryo. pull down pull down western blot western blot Immuno blot Immuno blot Gupta R., Mould RM., He Z., Luan S. sluan@nature.berkeley.edu. sluan@nature.berkeley.edu Accepted 2009-NOV-9 by ORCHARD. Arabidopsis ACT cDNA expression library CD4-22. Description: Generated from mRNA isolated from 3 day-old etiolated Columbia seedlings; library consists of 36,000,000 independent recombinants and was amplified on LE392; 22 of 23 clones tested contained inserts. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Only protein-protein interactions imex curation A chloroplast FKBP interacts with and affects the accumulation of Rieske subunit of cytochrome bf complex. A chloroplast FKBP interacts with and affects the accumulation of Rieske subunit of cytochrome bf complex. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 imex curation Gupta R., Mould RM., He Z., Luan S. sluan@nature.berkeley.edu Arabidopsis ACT cDNA expression library CD4-22. Description: Generated from mRNA isolated from 3 day-old etiolated Columbia seedlings; library consists of 36,000,000 independent recombinants and was amplified on LE392; 22 of 23 clones tested contained inserts. orchard 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 Gupta R., Mould RM., He Z., Luan S. sluan@nature.berkeley.edu. sluan@nature.berkeley.edu Accepted 2009-NOV-9 by ORCHARD. Arabidopsis ACT cDNA expression library CD4-22. Description: Generated from mRNA isolated from 3 day-old etiolated Columbia seedlings; library consists of 36,000,000 independent recombinants and was amplified on LE392; 22 of 23 clones tested contained inserts. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Only protein-protein interactions imex curation pnsl4_arath Photosynthetic NDH subunit of lumenal location 4, chloroplastic PNSL4 FK506-binding protein 16-2 FKBP16-2 FKBP22-2 FKBP23I Immunophilin FKBP16-2 Peptidyl-prolyl cis-trans isomerase FKBP16-2 Rotamase T19P19.100 At4g39710 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAISTLTLTQSLYTRSFRPTIFFSSSSSSSFSCLCSSSSDCEPKLSVKKRVFGVGLGFLASSILSLTPLDADATRIDYYATVGDPLCEYSYAKSGLGFCDLDVGFGDEAPRGVLVNIHYTARFADGTLFDSSYKRARPLTMRIGVGKVIRGLDQGILGGEGVPPMRVGGKRKLQIPPKLAYGPEPAGCFSGDCNIPGNATLLYDINFVEIYPGSNTR 1122EE1BAFD20E19 ucria_arath Cytochrome b6-f complex iron-sulfur subunit, chloroplastic petC PGR1 Rieske iron-sulfur protein Plastohydroquinone:plastocyanin oxidoreductase iron-sulfur protein Proton gradient regulation protein 1 F4C21.21 At4g03280 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MASSSLSPATQLGSSRSALMAMSSGLFVKPTKMNHQMVRKEKIGLRISCQASSIPADRVPDMEKRKTLNLLLLGALSLPTGYMLVPYATFFVPPGTGGGGGGTPAKDALGNDVVAAEWLKTHGPGDRTLTQGLKGDPTYLVVENDKTLATYGINAVCTHLGCVVPWNKAENKFLCPCHGSQYNAQGRVVRGPAPLSLALAHADIDEAGKVLFVPWVETDFRTGDAPWWS D79FAE03D85EA2F5 fkb23i-petc 1 4 unspecified role unspecified role bait bait 5 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 physical association physical association The precursor of AtFKBP13 was used as a bait in fusion with Gal4 DNA-binding domain to screen an Arabidopsis cDNA library in Gal4 activation-domain vector. Among the AtFKBP13 interacting clones that were sequenced, a number of them encoded the Rieske protein of various length fused in frame with the activation domain. The authors state that the longest isolated clone "had four amino acid truncations in the N terminus". This information was used for the amino acid range of the prey protein. fkb23i-petc-1 3 4 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 12 14 physical association physical association We conclude that the transit peptide of AtFKBP13 is required and, to certain degree, sufficient for the interaction with Rieske protein. The mature region, more specifically, the lumen domain, of Rieske protein is sufficient for interaction. Figure 3 petc-fkb23i 2 4 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 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 region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 17 20 physical association physical association As shown in Fig. ​Fig.4,4, precursor, mature, or lumen domain of Rieske protein copurified with the precursor but not with the mature form of AtFKBP13 Figure 4