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 novel interaction partner for the C-terminus of Arabidopsis thaliana plasma membrane H+-ATPase (AHA1 isoform): site and mechanism of action on H+-ATPase activity differ from those of 14-3-3 proteins. A novel interaction partner for the C-terminus of Arabidopsis thaliana plasma membrane H+-ATPase (AHA1 isoform): site and mechanism of action on H+-ATPase activity differ from those of 14-3-3 proteins. Plant J. (0960-7412) 2002 imex curation Morandini P., Valera M., Albumi C., Bonza MC., Giacometti S., Ravera G., Murgia I., Soave C., De Michelis MI. piero.morandini@unimi.it 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library provided by the Arabidopsis Biological Resource Center (ABRC) (cat.#CD4-10) 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 Morandini P., Valera M., Albumi C., Bonza MC., Giacometti S., Ravera G., Murgia I., Soave C., De Michelis MI. It was not possible to unambiguously identify proteins named as SNF1 and SNF4 in the paper. Therefore these proteins were not curated. piero.morandini@unimi.it Accepted 2009-JUL-23 by ORCHARD. 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library provided by the Arabidopsis Biological Resource Center (ABRC) (cat.#CD4-10) Plant J. (0960-7412) 2002 orchard Only protein-protein interactions imex curation A novel interaction partner for the C-terminus of Arabidopsis thaliana plasma membrane H+-ATPase (AHA1 isoform): site and mechanism of action on H+-ATPase activity differ from those of 14-3-3 proteins. A novel interaction partner for the C-terminus of Arabidopsis thaliana plasma membrane H+-ATPase (AHA1 isoform): site and mechanism of action on H+-ATPase activity differ from those of 14-3-3 proteins. Plant J. (0960-7412) 2002 imex curation Morandini P., Valera M., Albumi C., Bonza MC., Giacometti S., Ravera G., Murgia I., Soave C., De Michelis MI. piero.morandini@unimi.it 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library provided by the Arabidopsis Biological Resource Center (ABRC) (cat.#CD4-10) orchard Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting anti tag western anti tag western blot anti tag western Morandini P., Valera M., Albumi C., Bonza MC., Giacometti S., Ravera G., Murgia I., Soave C., De Michelis MI. piero.morandini@unimi.it Accepted 2009-JUL-23 by ORCHARD. 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library provided by the Arabidopsis Biological Resource Center (ABRC) (cat.#CD4-10) Plant J. (0960-7412) 2002 orchard Only protein-protein interactions imex curation A novel interaction partner for the C-terminus of Arabidopsis thaliana plasma membrane H+-ATPase (AHA1 isoform): site and mechanism of action on H+-ATPase activity differ from those of 14-3-3 proteins. A novel interaction partner for the C-terminus of Arabidopsis thaliana plasma membrane H+-ATPase (AHA1 isoform): site and mechanism of action on H+-ATPase activity differ from those of 14-3-3 proteins. Plant J. (0960-7412) 2002 imex curation Morandini P., Valera M., Albumi C., Bonza MC., Giacometti S., Ravera G., Murgia I., Soave C., De Michelis MI. piero.morandini@unimi.it 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library provided by the Arabidopsis Biological Resource Center (ABRC) (cat.#CD4-10) 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 Morandini P., Valera M., Albumi C., Bonza MC., Giacometti S., Ravera G., Murgia I., Soave C., De Michelis MI. piero.morandini@unimi.it Accepted 2009-JUL-23 by ORCHARD. 01-Sep-2009: Contacted by JYOTI. Arabidopsis cDNA library provided by the Arabidopsis Biological Resource Center (ABRC) (cat.#CD4-10) Plant J. (0960-7412) 2002 orchard Only protein-protein interactions imex curation pma1_arath ATPase 1, plasma membrane-type AHA1 Proton pump 1 F19F24.16 At2g18960 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSGLEDIKNETVDLEKIPIEEVFQQLKCTREGLTTQEGEDRIVIFGPNKLEEKKESKILKFLGFMWNPLSWVMEAAALMAIALANGDNRPPDWQDFVGIICLLVINSTISFIEENNAGNAAAALMAGLAPKTKVLRDGKWSEQEAAILVPGDIVSIKLGDIIPADARLLEGDPLKVDQSALTGESLPVTKHPGQEVFSGSTCKQGEIEAVVIATGVHTFFGKAAHLVDSTNQVGHFQKVLTSIGNFCICSIAIGIAIEIVVMYPIQHRKYRDGIDNLLVLLIGGIPIAMPTVLSVTMAIGSHRLSQQGAITKRMTAIEEMAGMDVLCSDKTGTLTLNKLSVDKNLVEVFCKGVEKDQVLLFAAMASRVENQDAIDAAMVGMLADPKEARAGIREVHFLPFNPVDKRTALTYIDSDGNWHRVSKGAPEQILDLANARPDLRKKVLSCIDKYAERGLRSLAVARQVVPEKTKESPGGPWEFVGLLPLFDPPRHDSAETIRRALNLGVNVKMITGDQLAIGKETGRRLGMGTNMYPSAALLGTDKDSNIASIPVEELIEKADGFAGVFPEHKYEIVKKLQERKHIVGMTGDGVNDAPALKKADIGIAVADATDAARGASDIVLTEPGLSVIISAVLTSRAIFQRMKNYTIYAVSITIRIVFGFMLIALIWEFDFSAFMVLIIAILNDGTIMTISKDRVKPSPTPDSWKLKEIFATGIVLGGYQAIMSVIFFWAAHKTDFFSDKFGVRSIRDNNDELMGAVYLQVSIISQALIFVTRSRSWSFVERPGALLMIAFVIAQLVATLIAVYADWTFAKVKGIGWGWAGVIWIYSIVTYFPQDILKFAIRYILSGKAWASLFDNRTAFTTKKDYGIGEREAQWAQAQRTLHGLQPKEDVNIFPEKGSYRELSEIAEQAKRRAEIARLRELHTLKGHVESVAKLKGLDIDTAGHHYTV A613FF6070ECC9F3 ppi1_arath Proton pump-interactor 1 PPI1 F27G19.100 At4g27500 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGVEVVNSGGFEVAPAPFEGKPEKNGKLDQGKGDDAPINFGSVGELPKNAEENNNKVVNSDAPKNAAEEWPVAKQIHSFYLVKYRSYADPKIKAKLDLADKELEKLNKARTGVLDKLRAKRAERSELFDLLDPLKSERKGFNTMFDEKRKEMEPLQQALGKLRSNDGGSARGPAICSSEEELNSMIYSYQYRIQHESIPLTEEKQILKEIRLLEGTRDKVIANAAMRAKIKESMGQKDDIQGQVKLMGAGLDGVKKERQAISARINELSEKLKATKDEITVLENELKTVSEKRDKAYSNIHDLRRQRDETNSEYYQNRTVLNKARDLAAQKNISELEALANAEVEKFISLWCSKKNFREDYEKRILQSLDSRQLSRDGRMRNPDEKPLIAPEAAPSKATPSETEVVPKAKAKPQPKEEPVSAPKPDATVAQNTEKAKDAVKVKNVADDDDDEVYGLGKPQKEEKPVDAATAKEMRKQEEIAKAKQAMERKKKLAEKAAAKAAIRAQKEAEKKEKKEQEKKAKKKTGGNTETETEEVPEASEEEIEAPVQEEKPQKEKVFKEKPIRNRTRGRGPETIPRAILKRKKSTNYWVYAAPAALVVLLLLVLGYYYVL F0882E8AB50D089B aha1-ppi 3 4 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 5 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 physical association physical association Figure 2 The authors state that "several yeast clones isolated in independent transformations contained the same prey, coding for a 102 aa peptide, .However, later in the paper they state "the Ppi1 clone encoding the firstt 88 aa of the protein was recovered in a two-hybrid screen using the isolated C-terminus of isoform AHA1 of the PM H+-ATPase as bait".It is unclear when this latter example was recovered so I have used the longer peptide in the entry. aha1-ppi-1 1 4 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 5 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 physical association physical association Figure 1 The interaction was retained when the AHA1and PPI1 coding fragments were exchanged on the bait and prey plasmids (Figure 1). aha1-2 1 4 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 4 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 18 20 physical association physical association Figure 1 A new, unexpected interaction of AHA1 C-terminal tail with itself was revealed by the assay, suggesting that the last 103 aa may contain a homodimerization domain for the H+-ATPase aha1-ppi-2 2 5 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 region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role bait bait arath-cell-culture Arabidopsis thaliana cell culture plant_cell_culture plant cellular culture The culture conditions should always be cited in the experimental modifications annotation Either whole plant or undifferentiated plant cells maintained in culture. direct interaction direct interaction Figure 4 PPI1 binds to native and immunoprecipitated H+-ATPase. aha1-ppi-3 2 5 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 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 4 unspecified role unspecified role bait bait arath-cell-culture Arabidopsis thaliana cell culture plant_cell_culture plant cellular culture The culture conditions should always be cited in the experimental modifications annotation Either whole plant or undifferentiated plant cells maintained in culture. direct interaction direct interaction Figure 4 PPI1 binds to native and immunoprecipitated H+-ATPase. aha1-ppi-6 1 5 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 4 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 35 33 physical association physical association Figure 1