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/ SARAF Inactivates the Store Operated Calcium Entry Machinery to Prevent Excess Calcium Refilling. SARAF Inactivates the Store Operated Calcium Entry Machinery to Prevent Excess Calcium Refilling. Cell (0092-8674) 2012 imex curation Palty R., Raveh A., Kaminsky I., Meller R., Reuveny E. Source of STIM1 and ORA1 taken from previous work of contributing scientist (PMID: 16906149, 18995841) e.reuveny@weizmann.ac.il 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Palty R., Raveh A., Kaminsky I., Meller R., Reuveny E. Source of STIM1 and ORA1 taken from previous work of contributing scientist (PMID: 16906149, 18995841) e.reuveny@weizmann.ac.il Accepted 2012-MAY-21 by JYOTI 20-Jun-2012: Contacted by JYOTI. Cell (0092-8674) 2012 Only protein-protein interactions imex curation SARAF Inactivates the Store Operated Calcium Entry Machinery to Prevent Excess Calcium Refilling. SARAF Inactivates the Store Operated Calcium Entry Machinery to Prevent Excess Calcium Refilling. Cell (0092-8674) 2012 imex curation Palty R., Raveh A., Kaminsky I., Meller R., Reuveny E. Source of STIM1 and ORA1 taken from previous work of contributing scientist (PMID: 16906149, 18995841) e.reuveny@weizmann.ac.il 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Palty R., Raveh A., Kaminsky I., Meller R., Reuveny E. S5A requires an isoform not yet in Swiss-Prot. I've requested that it be added, bit as its only a colocalisation, have put the entry for checking and release as it seems a bit silly to wait 3 months just for that. I'll add it when SP finally get the entry updated and public. Source of STIM1 and ORA1 taken from previous work of contributing scientist (PMID: 16906149, 18995841) e.reuveny@weizmann.ac.il Accepted 2012-MAY-21 by JYOTI Cell (0092-8674) 2012 Only protein-protein interactions imex curation SARAF Inactivates the Store Operated Calcium Entry Machinery to Prevent Excess Calcium Refilling. SARAF Inactivates the Store Operated Calcium Entry Machinery to Prevent Excess Calcium Refilling. Cell (0092-8674) 2012 imex curation Palty R., Raveh A., Kaminsky I., Meller R., Reuveny E. Source of STIM1 and ORA1 taken from previous work of contributing scientist (PMID: 16906149, 18995841) e.reuveny@weizmann.ac.il 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Palty R., Raveh A., Kaminsky I., Meller R., Reuveny E. Source of STIM1 and ORA1 taken from previous work of contributing scientist (PMID: 16906149, 18995841) e.reuveny@weizmann.ac.il Accepted 2012-MAY-21 by JYOTI 20-Jun-2012: Contacted by JYOTI. Cell (0092-8674) 2012 Only protein-protein interactions imex curation saraf_human Store-operated calcium entry-associated regulatory factor SARAF HBV X-transactivated gene 3 protein HBV XAg-transactivated protein 3 Protein FOAP-7 TMEM66 Transmembrane protein 66 XTP3 HSPC035 NPD003 PSEC0019 UNQ1967/PRO4499 protein protein human Homo sapiens Human MAAACGPGAAGYCLLLGLHLFLLTAGPALGWNDPDRMLLRDVKALTLHYDRYTTSRRLDPIPQLKCVGGTAGCDSYTPKVIQCQNKGWDGYDVQWECKTDLDIAYKFGKTVVSCEGYESSEDQYVLRGSCGLEYNLDYTELGLQKLKESGKQHGFASFSDYYYKWSSADSCNMSGLITIVVLLGIAFVVYKLFLSDGQYSPPPYSEYPPFSHRYQRFTNSAGPPPPGFKSEFTGPQNTGHGATSGFGSAFTGQQGYENSGPGFWTGLGTGGILGYLFGSNRAATPFSDSWYYPSYPPSYPGTWNRAYSPLHGGSGSYSVCSNSDTKTRTASGYGGTRRR 83CC0517AB635FE9 stim1_human Stromal interaction molecule 1 STIM1 GOK protein protein human Homo sapiens Human MDVCVRLALWLLWGLLLHQGQSLSHSHSEKATGTSSGANSEESTAAEFCRIDKPLCHSEDEKLSFEAVRNIHKLMDDDANGDVDVEESDEFLREDLNYHDPTVKHSTFHGEDKLISVEDLWKAWKSSEVYNWTVDEVVQWLITYVELPQYEETFRKLQLSGHAMPRLAVTNTTMTGTVLKMTDRSHRQKLQLKALDTVLFGPPLLTRHNHLKDFMLVVSIVIGVGGCWFAYIQNRYSKEHMKKMMKDLEGLHRAEQSLHDLQERLHKAQEEHRTVEVEKVHLEKKLRDEINLAKQEAQRLKELREGTENERSRQKYAEEELEQVREALRKAEKELESHSSWYAPEALQKWLQLTHEVEVQYYNIKKQNAEKQLLVAKEGAEKIKKKRNTLFGTFHVAHSSSLDDVDHKILTAKQALSEVTAALRERLHRWQQIEILCGFQIVNNPGIHSLVAALNIDPSWMGSTRPNPAHFIMTDDVDDMDEEIVSPLSMQSPSLQSSVRQRLTEPQHGLGSQRDLTHSDSESSLHMSDRQRVAPKPPQMSRAADEALNAMTSNGSHRLIEGVHPGSLVEKLPDSPALAKKALLALNHGLDKAHSLMELSPSAPPGGSPHLDSSRSHSPSSPDPDTPSPVGDSRALQASRNTRIPHLAGKKAVAEEDNGSIGEETDSSPGRKKFPLKIFKKPLKK DIP protein Q13586 original sequence version: 141 DIP protein Q13586 original sequence version: 99 DIP protein Q13586 original sequence version: 108 DIP protein Q13586 original sequence version: 121 DIP protein Q13586 original sequence version: 125 DIP protein Q13586 original sequence version: 100 FFB5E7CB3D68A7A6 crcm1_human Calcium release-activated calcium channel protein 1 ORAI1 CRACM1 TMEM142A Protein orai-1 Transmembrane protein 142A protein protein human Homo sapiens Human MHPEPAPPPSRSSPELPPSGGSTTSGSRRSRRRSGDGEPPGAPPPPPSAVTYPDWIGQSYSEVMSLNEHSMQALSWRKLYLSRAKLKASSRTSALLSGFAMVAMVEVQLDADHDYPPGLLIAFSACTTVLVAVHLFALMISTCILPNIEAVSNVHNLNSVKESPHERMHRHIELAWAFSTVIGTLLFLAEVVLLCWVKFLPLKKQPGQPRPTSKPPASGAAANVSTSGITPGQAAAIASTTIMVPFGLIFIVFAVHFYRSLVSHKTDRQFQELNELAEFARLQDQLDHRGDHPLTPGSHYA DIP protein Q96D31 original sequence version: 95 DIP protein Q96D31 original sequence version: 38 DIP protein Q96D31 original sequence version: 58 DIP protein Q96D31 original sequence version: 55 DIP protein Q96D31 original sequence version: 57 DIP protein Q96D31 original sequence version: 53 DIP protein Q96D31 original sequence version: 63 DIP protein Q96D31 original sequence version: 76 DIP protein Q96D31 original sequence version: 79 DIP protein Q96D31 original sequence version: 56 F85F7C06F36628D4 stim1-tmem66 2 4 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role neutral component neutral component region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Fig. 1G, 3B, Supp Fig. 5B and Movie S1 Addition of Thapsigargin (an irreversible sarcoplasmic/endoplasmic reticulum Ca2+-ATPase), induced translocation of the complex to the plasma membrane. orai1-tmem66 2 4 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role neutral component neutral component region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Fig. 3C, 3D, Supp. Fig. 4D Silencing STIM1 and STIM2 protein levels inhibited the Tg-induced SARAF-GFP translocation in the presence of Orai1-mCherry tmem66-stim1 3 4 unspecified role unspecified role bait bait region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 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 physical association physical association Fig. 5C, 5D Traces of Orai1 immunopercipitate were also detected, which the authors attributed to its native interaction with STIM1 orai1-stim1-8 1 4 unspecified role unspecified role fluorescence donor fluorescence donor region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 ancillary ancillary unspecified role unspecified role proximity proximity close-contact colocalization neighbourhood interaction Fig. 4C, 5D, Supp. Fig. 6 results indicate that SARAF and STIM1 associate under resting conditions and that store operated calcium entry activation promotes molecular rearrangements between STIM1 and SARAF C termini in an Orai1-dependent manner. orai1-stim1-7 2 6 unspecified role unspecified role neutral component neutral component region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Thapsigargin (an irreversible sarcoplasmic/endoplasmic reticulum Ca2+-ATPase) Fig. 6A