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/ park-2009-4 STIM1 clusters and activates CRAC channels via direct binding of a cytosolic domain to Orai1. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by comassie molecular weight estimation by coomasie staining weight by comassie Park CY., Hoover PJ., Mullins FM., Bachhawat P., Covington ED., Raunser S., Walz T., Garcia KC., Dolmetsch RE., Lewis RS. Purified proteins were fractionated on a Superose 6 gel filtration column. Molecular weight determination of the STIM1 tetramer was confrimed by multiangle light scattering (MALS). Source of STIM1 taken from PMID: 16966423 and of ORAI1 from the work of the contributing author described in PMID:17293345 ricardo.dolmetsch@stanford.edu, rslewis@stanford.edu Cell (0092-8674) 2009 Only protein-protein interactions 8 imex curation park-2009-1 STIM1 clusters and activates CRAC channels via direct binding of a cytosolic domain to Orai1. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Park CY., Hoover PJ., Mullins FM., Bachhawat P., Covington ED., Raunser S., Walz T., Garcia KC., Dolmetsch RE., Lewis RS. Source of STIM1 taken from PMID: 16966423 and of ORAI1 from the work of the contributing author described in PMID:17293345 ricardo.dolmetsch@stanford.edu, rslewis@stanford.edu Cell (0092-8674) 2009 Only protein-protein interactions 8 imex curation park-2009-2 STIM1 clusters and activates CRAC channels via direct binding of a cytosolic domain to Orai1. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast ub reconstruction ubiquitin reconstruction ub reconstruction predetermined predetermined participant predetermined Park CY., Hoover PJ., Mullins FM., Bachhawat P., Covington ED., Raunser S., Walz T., Garcia KC., Dolmetsch RE., Lewis RS. Source of STIM1 taken from PMID: 16966423 and of ORAI1 from the work of the contributing author described in PMID:17293345 ricardo.dolmetsch@stanford.edu, rslewis@stanford.edu Cell (0092-8674) 2009 Only protein-protein interactions 8 imex curation park-2009-3 STIM1 clusters and activates CRAC channels via direct binding of a cytosolic domain to Orai1. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Park CY., Hoover PJ., Mullins FM., Bachhawat P., Covington ED., Raunser S., Walz T., Garcia KC., Dolmetsch RE., Lewis RS. Source of STIM1 taken from PMID: 16966423 and of ORAI1 from the work of the contributing author described in PMID:17293345 ricardo.dolmetsch@stanford.edu, rslewis@stanford.edu Cell (0092-8674) 2009 Only protein-protein interactions 8 imex curation park-2009-5 STIM1 clusters and activates CRAC channels via direct binding of a cytosolic domain to Orai1. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Park CY., Hoover PJ., Mullins FM., Bachhawat P., Covington ED., Raunser S., Walz T., Garcia KC., Dolmetsch RE., Lewis RS. Source of STIM1 taken from PMID: 16966423 and of ORAI1 from the work of the contributing author described in PMID:17293345 ricardo.dolmetsch@stanford.edu, rslewis@stanford.edu Cell (0092-8674) 2009 Only protein-protein interactions 8 imex curation 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 stim1_human Stromal interaction molecule 1 STIM1 GOK protein protein human Homo sapiens Human MDVCVRLALWLLWGLLLHQGQSLSHSHSEKATGTSSGANSEESTAAEFCRIDKPLCHSEDEKLSFEAVRNIHKLMDDDANGDVDVEESDEFLREDLNYHDPTVKHSTFHGEDKLISVEDLWKAWKSSEVYNWTVDEVVQWLITYVELPQYEETFRKLQLSGHAMPRLAVTNTTMTGTVLKMTDRSHRQKLQLKALDTVLFGPPLLTRHNHLKDFMLVVSIVIGVGGCWFAYIQNRYSKEHMKKMMKDLEGLHRAEQSLHDLQERLHKAQEEHRTVEVEKVHLEKKLRDEINLAKQEAQRLKELREGTENERSRQKYAEEELEQVREALRKAEKELESHSSWYAPEALQKWLQLTHEVEVQYYNIKKQNAEKQLLVAKEGAEKIKKKRNTLFGTFHVAHSSSLDDVDHKILTAKQALSEVTAALRERLHRWQQIEILCGFQIVNNPGIHSLVAALNIDPSWMGSTRPNPAHFIMTDDVDDMDEEIVSPLSMQSPSLQSSVRQRLTEPQHGLGSQRDLTHSDSESSLHMSDRQRVAPKPPQMSRAADEALNAMTSNGSHRLIEGVHPGSLVEKLPDSPALAKKALLALNHGLDKAHSLMELSPSAPPGGSPHLDSSRSHSPSSPDPDTPSPVGDSRALQASRNTRIPHLAGKKAVAEEDNGSIGEETDSSPGRKKFPLKIFKKPLKK orai1-5 Homodimerisation of ORAI1 801564 n/a 801569 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n-terminal EEYMPME tag tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 4C stim1-5 Homotetramerisation of STIM1 801564 n/a 801570 unspecified role unspecified role neutral component neutral component cad region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false Stoichiometry: 4.0 direct interaction direct interaction false false false Fig. 4D orai1-stim1-5 Interaction between ORAI1 and STIM1 801564 n/a 801569 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n-terminal EEYMPME tag tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. n/a 801570 unspecified role unspecified role neutral component neutral component cad region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. physical association physical association false false false Fig. 4C The complex was stable in 0.5 M NaCl, suggesting a high-affinity hydrophobic interaction of multiple CAD and Orai1 proteins. orai1-stim1-1 Interaction between ORAI1 and STIM1 identified by coimmunoprecipitation 801565 n/a 801570 unspecified role unspecified role prey prey cad region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false n/a 801569 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 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 3B, Supp. Fig. 8 orai1-stim1-2 Interaction between ORAI1 and STIM1 identified by coimmunoprecipitation 801565 n/a 801570 unspecified role unspecified role bait bait cad region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false n/a 801569 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 n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 3C, 5B, 5D orai1-stim1-3 Interaction between ORAI1 and STIM1 identified by ubiquitin reconstruction 801566 n/a 801570 unspecified role unspecified role bait bait cad region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 801569 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 3F, 5A, 5C orai1-stim1-4 Interaction between ORAI1 and STIM1 identified by pulldown 801567 n/a 801570 unspecified role unspecified role bait bait cad region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) n/a 801569 unspecified role unspecified role prey prey c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n-terminal EEYMPME tag tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. physical association physical association false false false Fig. 4A orai1-stim1-6 Colocalisation of ORAI1 and STIM1 801568 n/a 801569 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 801570 unspecified role unspecified role neutral component neutral component region rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. 1C