MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it http://mint.bio.uniroma2.it/mint anonymous-2014k-1 Atomic force microscopy (AFM) imaging suggests that stromal interaction molecule 1 (STIM1) binds to Orai1 with six-fold symmetry in vitro In vitro atomic force microsc atomic force microscopy AFM AFM atomic force microsc predetermined predetermined participant predetermined jme1000@cam.ac.uk 2014 imex curation Balasuriya D., Srivats S., Murrell-Lagnado R.D., Michael Edwardson J. FEBS Lett. (0014-5793) Only protein-protein interactions anonymous-2014e-2 Atomic force microscopy (AFM) imaging suggests that stromal interaction molecule 1 (STIM1) binds to Orai1 with six-fold symmetry human-tsa201 Homo sapiens subclone of human embryonic kidney 293 cell line that expresses simian virus 40 T antigen tsa201 Human embryonic kidney cell line subclone of 293 cell line that expresses simian virus 40 T antigen fluorescence imaging fluorescence microscopy fluorescence imaging anti tag immunost anti tag immunostaining anti tag immunost jme1000@cam.ac.uk 2014 imex curation Balasuriya D., Srivats S., Murrell-Lagnado R.D., Michael Edwardson J. FEBS Lett. (0014-5793) Only protein-protein interactions anonymous-2014y-4 Atomic force microscopy (AFM) imaging suggests that stromal interaction molecule 1 (STIM1) binds to Orai1 with six-fold symmetry human-tsa201 Homo sapiens subclone of human embryonic kidney 293 cell line that expresses simian virus 40 T antigen tsa201 Human embryonic kidney cell line subclone of 293 cell line that expresses simian virus 40 T antigen anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western jme1000@cam.ac.uk 2014 imex curation Balasuriya D., Srivats S., Murrell-Lagnado R.D., Michael Edwardson J. FEBS Lett. (0014-5793) Only protein-protein interactions 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 orai1-1 1548833 n/a Orai1 1548836 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tag 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: 6.0 direct interaction direct interaction false false false f1 orai1-stim1-13 1548833 n/a STIM1 1548837 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a Orai1 1548836 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his/myc tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f3b f3c f3d f3f f3g stim1-1 1548833 n/a STIM1 1548837 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction false false false f4 A low-magnification AFM image of the isolated protein (Fig. 4B) shows single small particles, as well as doublets (arrowheads), and string-like structures (arrow) similar to those described above. Zoomed images of these two types of multimeric structures are shown in Fig. 4C. A frequency distribution of the volumes of the STIM1 particles (Fig. 4D) stim1-stim1-1 1548833 n/a STIM1 1548837 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a STIM1 1548837 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction false false false f4 A low-magnification AFM image of the isolated protein (Fig. 4B) shows single small particles, as well as doublets (arrowheads), and string-like structures (arrow) similar to those described above. Zoomed images of these two types of multimeric structures are shown in Fig. 4C. A frequency distribution of the volumes of the STIM1 particles (Fig. 4D) orai1-stim1-10 1548834 n/a Orai1 1548836 anti tag immunost anti tag immunostaining anti tag immunost unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell myc/his tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a STIM1 1548837 anti tag immunost anti tag immunostaining anti tag immunost unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false f2a Cells were next co-transfected with DNA encoding Orai1-Myc/His and HA-STIM1. The localization of the two proteins, with and without thapsigargin (5 M) treatment, was determined by immunofluorescence analysis of saponin-permeabilized cells. In control cells, Orai1 was seen predominantly at the plasma membrane, whereas STIM1 was found in a reticular pattern throughout the cell cytoplasm, consistent with an ER localization (Fig. 2A). By contrast, in thapsigargin-treated cells, the STIM1 and Orai1 signals overlapped extensively, consistent with the translocation of STIM1 to sites beneath the plasma membrane orai1-stim1-12 1548835 n/a Orai1 1548836 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell myc/his tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a STIM1 1548837 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f2b f3a Co-transfected cells, either untreated or thapsigargin-treated, were solubilized, and the detergent extracts were subjected to anti-Myc immunoprecipitation followed by immunoblotting with anti-Myc (Orai1) and anti-HA (STIM1) antibodies. Orai1 was present in both samples, as expected (Fig. 2B), whereas STIM1 was coimmunopreciptated only from the extract of thapsigargin-treated cells, indicating that STIM1 associates with Orai1 only after store depletion.