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/ wang-2008-2 Characterization of the Arabidopsis heterotrimeric G protein. arath-leaf Arabidopsis thaliana leaf leaf leaf fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Wang S., Assmann SM., Fedoroff NV. The authors confirmed the identities of the proteins used in this study. Transformed Arabidopsis mesophyll protoplasts used for in vivo FRET. nvf1@psu.edu J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation wang-2008-1 Characterization of the Arabidopsis heterotrimeric G protein. arath-leaf Arabidopsis thaliana leaf leaf leaf confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Wang S., Assmann SM., Fedoroff NV. The authors confirmed the identities of the proteins used in this study. nvf1@psu.edu J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation wang-2008-3 Characterization of the Arabidopsis heterotrimeric G protein. arath-leaf Arabidopsis thaliana leaf leaf leaf anti bait coip anti bait coimmunoprecipitation anti bait coip weight identificat confirmation by molecular weight weight identificat Wang S., Assmann SM., Fedoroff NV. As the anti-body used in western blotting was not specific , mouse monoclonal anti-G/C/Y antibodies (anti-G/C/Y antibody detects GFP, CFP and YFP proteins)), protein indentification was made on the basis of protein size using electrophoresis through a discontinuous polyacrylamide gel. The authors confirmed the identities of the proteins used in this study. Transformed Arabidopsis mesophyll protoplasts used. nvf1@psu.edu J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation gbb_arath Guanine nucleotide-binding protein subunit beta GB1 T4L20.40 At4g34460 transducin AGB1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSVSELKERHAVATETVNNLRDQLRQRRLQLLDTDVARYSAAQGRTRVSFGATDLVCCRTLQGHTGKVYSLDWTPERNRIVSASQDGRLIVWNALTSQKTHAIKLPCAWVMTCAFSPNGQSVACGGLDSVCSIFSLSSTADKDGTVPVSRMLTGHRGYVSCCQYVPNEDAHLITSSGDQTCILWDVTTGLKTSVFGGEFQSGHTADVLSVSISGSNPNWFISGSCDSTARLWDTRAASRAVRTFHGHEGDVNTVKFFPDGYRFGTGSDDGTCRLYDIRTGHQLQVYQPHGDGENGPVTSIAFSVSGRLLFAGYASNNTCYVWDTLLGEVVLDLGLQQDSHRNRISCLGLSADGSALCTGSWDSNLKIWAFGGHRRVI gg1_arath Guanine nucleotide-binding protein subunit gamma 1 GG1 AGG1 Ggamma-subunit 1 Heterotrimeric G protein gamma-subunit 1 MAA21.50 At3g63420 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MREETVVYEQEESVSHGGGKHRILAELARVEQEVAFLEKELKEVENTDIVSTVCEELLSVIEKGPDPLLPLTNGPLNLGWDRWFEGPNGGEGCRCLIL gpa1_arath Guanine nucleotide-binding protein alpha-1 subunit GPA1 At2g26300 T1D16.6 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGLLCSRSRHHTEDTDENTQAAEIERRIEQEAKAEKHIRKLLLLGAGESGKSTIFKQIKLLFQTGFDEGELKSYVPVIHANVYQTIKLLHDGTKEFAQNETDSAKYMLSSESIAIGEKLSEIGGRLDYPRLTKDIAEGIETLWKDPAIQETCARGNELQVPDCTKYLMENLKRLSDINYIPTKEDVLYARVRTTGVVEIQFSPVGENKKSGEVYRLFDVGGQRNERRKWIHLFEGVTAVIFCAAISEYDQTLFEDEQKNRMMETKELFDWVLKQPCFEKTSFMLFLNKFDIFEKKVLDVPLNVCEWFRDYQPVSSGKQEIEHAYEFVKKKFEELYYQNTAPDRVDRVFKIYRTTALDQKLVKKTFKLVDETLRRRNLLEAGLL gb1-agg1-1 GB1 (At4g34460) interacts with AGG1 (At3g63420) 732467 N/A 732470 unspecified role unspecified role neutral component neutral component n-terminal cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 732471 unspecified role unspecified role neutral component neutral component n-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false direct interaction direct interaction false false false Figure 3A Colocalisation occurs in the plasma membrane. gb1-agg1-gpa1 GPA1 (At2g26300) forms a heterotrimer with GB1 (At4g34460) and AGG1 (At3g63420) 732467 N/A 732471 unspecified role unspecified role neutral component neutral component n-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 732470 unspecified role unspecified role neutral component neutral component n-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 732472 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag certain certain sequence position certain certain certain sequence position certain false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 3D Figure 3I The cfp tag was positioned between amino acid residues 97 and 98 in the GPA1 protein The authors state that "the G alpha subunit does not interact with either G beta or G gamma1 alone in protoplasts, but interacts with the G beta /gamma complex". P18064 over-expressed P49177 over-expressed Q9FDX9 over-expressed gb1-agg1-gpa1-1 GPA1 (At2g26300) forms a heterotrimer with GB1 (At4g34460) and AGG1 (At3g63420) 732467 N/A 732470 unspecified role unspecified role neutral component neutral component N/A 732471 unspecified role unspecified role neutral component neutral component n-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 732472 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag certain certain sequence position certain certain certain sequence position certain false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 3F Figure 3I The cfp tag was positioned between amino acid residues 97 and 98 in the GPA1 protein The authors concluded that "the G alpha subunit does not interact with either Gbeta or G gamma1 alone in protoplasts, but interacts with the G beta/gamma complex.". P18064 over-expressed P49177 over-expressed Q9FDX9 over-expressed gb1-agg1 GB1 (At4g34460) interacts with AGG1 (At3g63420) 732468 N/A 732470 unspecified role unspecified role neutral component neutral component n-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 732471 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Figure 2 Localisation of GB1 to the plasma membrane was dependent on the presence of AGG1. Q9FDX9 over-expressed gpa1-agg1 GPA1 (At2g26300) interacts with AGG1 (At3g63420) only in the presence of GB1 (At4g34460) 732469 N/A 732470 unspecified role unspecified role ancillary ancillary N/A 732471 unspecified role unspecified role prey prey n-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 732472 unspecified role unspecified role bait bait region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag certain certain sequence position certain certain certain sequence position certain false association association false false false Figure 4 The cfp tag was positioned between amino acid residues 97 and 98 in the GPA1 protein The authors state that " the subunits of the Arabidopsis G protein form a heterotrimeric complex in vivo and that the interaction of G alpha and G gamma 1 requires G beta". Q9FDX9 over-expressed P49177 over-expressed P18064 over-expressed Protein indentification was made on the basis of protein size using electrophoresis through a 12 percent discontinuous polyacrylamide gel.