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/ kato-2004-2 Characterization of heterotrimeric G protein complexes in rice plasma membrane. 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 Kato C., Mizutani T., Tamaki H., Kumagai H., Kamiya T., Hirobe A., Fujisawa Y., Kato H., Iwasaki Y. Beta galactosidase assays were carried out to ascertain the strength of protein interaction. The UniProt accession number for RGG2 was determined from the amino acid sequence given in Figure 1 of this paper using a amino acid similarity search against the UniProt database. iwasaki@fpu.ac.jp Plant J. (0960-7412) 2004 Only protein-protein interactions imex curation In some assays plama membranes from a mutant rice plants carrying a constitutively active form of the GPA1 protein (GPA1 Q223L) were analysed. kato-2004-3 Characterization of heterotrimeric G protein complexes in rice plasma membrane. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Kato C., Mizutani T., Tamaki H., Kumagai H., Kamiya T., Hirobe A., Fujisawa Y., Kato H., Iwasaki Y. The Uniprot accession number for RGG2 was determined using the amino acid sequence available in Figure 1 and an amino acid similarity search against the UniProt database. iwasaki@fpu.ac.jp Plant J. (0960-7412) 2004 Only protein-protein interactions imex curation In some assays plama membranes from a mutant rice plants carrying a constitutively active form of the GPA1 protein (GPA1 Q223L) were analysed. The UniProt accession number for RGG2 was determined from the amino acid sequence given in Figure 1 of this paper using a amino acid similarity search against the UniProt database. kato-2004-4 Characterization of heterotrimeric G protein complexes in rice plasma membrane. in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Kato C., Mizutani T., Tamaki H., Kumagai H., Kamiya T., Hirobe A., Fujisawa Y., Kato H., Iwasaki Y. The Uniprot accession number for RGG2 was determined using the amino acid sequence available in Figure 1 and an amino acid similarity search against the UniProt database. iwasaki@fpu.ac.jp Plant J. (0960-7412) 2004 Only protein-protein interactions imex curation In some assays plama membranes from a mutant rice plants carrying a constitutively active form of the GPA1 protein (GPA1 Q223L) were analysed. The UniProt accession number for RGG2 was determined from the amino acid sequence given in Figure 1 of this paper using a amino acid similarity search against the UniProt database. kato-2004-1 Characterization of heterotrimeric G protein complexes in rice plasma membrane. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column monoclonal western monoclonal antibody western blot monoclonal western Kato C., Mizutani T., Tamaki H., Kumagai H., Kamiya T., Hirobe A., Fujisawa Y., Kato H., Iwasaki Y. In some assays plama membranes from a mutant rice plants carrying a constitutively active form of the GPA1 protein (GPA1 Q223L) were analysed. The UniProt accession number for RGG2 was determined from the amino acid sequence given in Figure 1 of this paper using a amino acid similarity search against the UniProt database. iwasaki@fpu.ac.jp Plant J. (0960-7412) 2004 Only protein-protein interactions imex curation gpa1_orysj Guanine nucleotide-binding protein alpha-1 subunit GPA1 D1 GA1 RGA1 OJ1005_D04.15 OSJNBa0049D13.1 Protein Dwarf1 LOC_Os05g26890 Os05g0333200 Protein Daikoku dwarf protein protein orysj Oryza sativa subsp. japonica (Rice) Rice MGSSCSRSHSLSEAETTKNAKSADIDRRILQETKAEQHIHKLLLLGAGESGKSTIFKQIKLLFQTGFDEAELRSYTSVIHANVYQTIKILYEGAKELSQVESDSSKYVISPDNQEIGEKLSDIDGRLDYPLLNKELVLDVKRLWQDPAIQETYLRGSILQLPDCAQYFMENLDRLAEAGYVPTKEDVLYARVRTNGVVQIQFSPVGENKRGGEVYRLYDVGGQRNERRKWIHLFEGVNAVIFCAAISEYDQMLFEDETKNRMMETKELFDWVLKQRCFEKTSFILFLNKFDIFEKKIQKVPLSVCEWFKDYQPIAPGKQEVEHAYEFVKKKFEELYFQSSKPDRVDRVFKIYRTTALDQKLVKKTFKLIDESMRRSREGT gbb_orysj Guanine nucleotide-binding protein subunit beta RGB1 WD40 repeat-containing protein 80 OSJNBa0039O18.9 LOC_Os03g46650 Os03g0669200 protein protein orysj Oryza sativa subsp. japonica (Rice) Rice MASVAELKEKHAAATASVNSLRERLRQRRQMLLDTDVERYSRTQGRTPVSFNPTDLVCCRTLQGHSGKVYSLDWTPEKNWIVSASQDGRLIVWNALTSQKTHAIKLHCPWVMTCAFAPNGQSVACGGLDSACSIFNLNSQADRDGNIPVSRILTGHKGYVSSCQYVPDQETRLITSSGDQTCVLWDVTTGQRISIFGGEFPSGHTADVLSLSINSSNSNMFVSGSCDATVRLWDIRIASRAVRTYHGHEGDINSVKFFPDGQRFGTGSDDGTCRLFDVRTGHQLQVYSREPDRNDNELPTVTSIAFSISGRLLFAGYSNGDCYVWDTLLAEVVLNLGNLQNSHEGRISCLGLSSDGSALCTGSWDKNLKIWAFSGHRKIV gg2_orysj Guanine nucleotide-binding protein subunit gamma 2 RGG2 Ggamma-subunit 2 Heterotrimeric G protein gamma-subunit 2 OSJNBa0026E05.11-1 LOC_Os02g04520 Os02g0137800 protein protein orysj Oryza sativa subsp. japonica (Rice) Rice MRGEANGEEEQQPPRRNHLRDDAEEEEEVERRAARPVSGQQQQQQRRRPTDVGGGAAMRSVGYVGKHRLSAAIARLDQELQSLQDELNELETMEPASAACQGVITSTEGKSDPLLPVTIGPENASWERWFQRVRSSRSNKWWASKGSDFS gg1_orysj Guanine nucleotide-binding protein subunit gamma 1 RGG1 Ggamma-subunit 1 Heterotrimeric G protein gamma-subunit 1 OsJ_11830 LOC_Os03g43480 Os03g0635100 protein protein orysj Oryza sativa subsp. japonica (Rice) Rice MQAGGGGDAGDTRGRHRIQAELKKLEQEARFLEEELEELDKTDKVSAALQELMVTAESKADPLLPVTTGPACQSWDRWFEGPQDLRRCKCWFL gbb-gpa1 GPA1 interacts with GBB 541668 N/A 541672 unspecified role unspecified role prey prey Q0DJ33:p.Gln223Leu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false N/A 541673 unspecified role unspecified role bait bait physical association physical association false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. Interaction between GBB and GPA1 was 'weak' , measured beta galactosidase activity was 25.7E-3 (GPA1 autoactivation measured beta galactosidase activity was 9.5E-3). Table 1 Interaction between GBB and GPA1 was 'weak' , measured beta galactosidase activity was 25.7E-3 (GPA1 autoactivation measured beta galactosidase activity was 9.5E-3). gbb-rrg2 RRG2 interacts with GBB 541668 N/A 541674 unspecified role unspecified role prey prey n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 541673 unspecified role unspecified role bait bait physical association physical association false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. Interaction between GBB and GPA1 was 'strong' , measured beta galactosidase activity was 13.2 Units(GPA1 autoactivation measured beta galactosidase activity was 6.9E-3 Units). Table 1 Interaction between GBB and GPA1 was 'strong' , measured beta galactosidase activity was 13.2 Units(GPA1 autoactivation measured beta galactosidase activity was 6.9E-3 Units). gbb-rrg1 RRG1 interacts with GBB 541668 N/A 541675 unspecified role unspecified role prey prey N/A 541673 unspecified role unspecified role bait bait physical association physical association false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. Interaction between GBB and GPA1 was 'strong' , measured beta galactosidase activity was 4.2 Units (GPA1 autoactivation measured beta galactosidase activity was 5.1E-3 Units). Table 1 Interaction between GBB and GPA1 was 'strong' , measured beta galactosidase activity was 4.2 Units (GPA1 autoactivation measured beta galactosidase activity was 5.1E-3 Units). gbb-rgg1-1 RGG1 interacts with GBB 541669 N/A 541675 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 541673 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 false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro physical association physical association false false false 5b Both proteins were labelled with 35S-methionine and 35S-cysteine gbb-rgg2-1 RGG2 interacts with GBB 541669 N/A 541673 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 false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 541674 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro physical association physical association false false false 5b Both proteins were labelled with 35S-methionine and 35S-cysteine gbb-rgg1-2 RGG1 interacts with GBB 541670 N/A 541673 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 541675 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro physical association physical association false false false 5a 5b Both proteins were labelled with 35S-methionine and 35S-cysteine gbb-rgg2-2 RGG2 interacts with GBB 541670 N/A 541674 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 541673 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro physical association physical association false false false 5a 5b Both proteins were labelled with 35S-methionine and 35S-cysteine rrg1-rrg2 The G protein complex 541671 N/A 541673 unspecified role unspecified role neutral component neutral component orysj Oryza sativa subsp. japonica (Rice) Rice N/A 541672 unspecified role unspecified role neutral component neutral component orysj Oryza sativa subsp. japonica (Rice) Rice N/A 541675 unspecified role unspecified role neutral component neutral component orysj Oryza sativa subsp. japonica (Rice) Rice N/A 541674 unspecified role unspecified role neutral component neutral component orysj Oryza sativa subsp. japonica (Rice) Rice physical association physical association false false false 4b 6e 7e Only an active form of GPA1 was seen in a complex of 60Kda made up of the RRG1, RGG2 and GBB proteins. A 400Kda protein complex containing RRG1, RRG2 , GBB and GPA1 was also observed in this assay oly when an inactive form of GPA1 was present.