UniProt UniProt http://www.uniprot.org BRI2 interacts with BACE1 and regulates its cellular levels by promoting its degradation and reducing its mRNA levels. BRI2 interacts with BACE1 and regulates its cellular levels by promoting its degradation and reducing its mRNA levels. Current Alzheimer research 2013 imex curation Tsachaki M., Fotinopoulou A., Slavi N., Zarkou V., Ghiso J., Efthimiopoulos S. efthis@biol.uoa.gr 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-sh_sy5y Homo sapiens SH-SY5Y cell line sh_sy5y Human neuroblastoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tsachaki M., Fotinopoulou A., Slavi N., Zarkou V., Ghiso J., Efthimiopoulos S. efthis@biol.uoa.gr Accepted 2019-MAY-13 AT 09:30 BST AT 09:30 BST by MGT Current Alzheimer research 2013 imex curation BRI2 interacts with BACE1 and regulates its cellular levels by promoting its degradation and reducing its mRNA levels. BRI2 interacts with BACE1 and regulates its cellular levels by promoting its degradation and reducing its mRNA levels. Current Alzheimer research 2013 imex curation Tsachaki M., Fotinopoulou A., Slavi N., Zarkou V., Ghiso J., Efthimiopoulos S. efthis@biol.uoa.gr 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Tsachaki M., Fotinopoulou A., Slavi N., Zarkou V., Ghiso J., Efthimiopoulos S. efthis@biol.uoa.gr Accepted 2019-MAY-13 AT 09:30 BST AT 09:30 BST by MGT Current Alzheimer research 2013 imex curation BRI2 interacts with BACE1 and regulates its cellular levels by promoting its degradation and reducing its mRNA levels. BRI2 interacts with BACE1 and regulates its cellular levels by promoting its degradation and reducing its mRNA levels. Current Alzheimer research 2013 imex curation Tsachaki M., Fotinopoulou A., Slavi N., Zarkou V., Ghiso J., Efthimiopoulos S. efthis@biol.uoa.gr 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 western blot western blot Immuno blot Immuno blot Tsachaki M., Fotinopoulou A., Slavi N., Zarkou V., Ghiso J., Efthimiopoulos S. efthis@biol.uoa.gr Accepted 2019-MAY-13 AT 09:30 BST AT 09:30 BST by MGT Current Alzheimer research 2013 imex curation BRI2 interacts with BACE1 and regulates its cellular levels by promoting its degradation and reducing its mRNA levels. BRI2 interacts with BACE1 and regulates its cellular levels by promoting its degradation and reducing its mRNA levels. Current Alzheimer research 2013 imex curation Tsachaki M., Fotinopoulou A., Slavi N., Zarkou V., Ghiso J., Efthimiopoulos S. efthis@biol.uoa.gr 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 immunocytochemistry immunocytochemistry Tsachaki M., Fotinopoulou A., Slavi N., Zarkou V., Ghiso J., Efthimiopoulos S. efthis@biol.uoa.gr Accepted 2019-MAY-13 AT 09:30 BST AT 09:30 BST by MGT Current Alzheimer research 2013 imex curation bace1_human Beta-secretase 1 BACE KIAA1149 Beta-site amyloid precursor protein cleaving enzyme 1 Membrane-associated aspartic protease 2 Memapsin-2 Aspartyl protease 2 BACE1 protein protein human Homo sapiens Human MAQALPWLLLWMGAGVLPAHGTQHGIRLPLRSGLGGAPLGLRLPRETDEEPEEPGRRGSFVEMVDNLRGKSGQGYYVEMTVGSPPQTLNILVDTGSSNFAVGAAPHPFLHRYYQRQLSSTYRDLRKGVYVPYTQGKWEGELGTDLVSIPHGPNVTVRANIAAITESDKFFINGSNWEGILGLAYAEIARPDDSLEPFFDSLVKQTHVPNLFSLQLCGAGFPLNQSEVLASVGGSMIIGGIDHSLYTGSLWYTPIRREWYYEVIIVRVEINGQDLKMDCKEYNYDKSIVDSGTTNLRLPKKVFEAAVKSIKAASSTEKFPDGFWLGEQLVCWQAGTTPWNIFPVISLYLMGEVTNQSFRITILPQQYLRPVEDVATSQDDCYKFAISQSSTGTVMGAVIMEGFYVVFDRARKRIGFAVSACHVHDEFRTAAVEGPFVTLDMEDCGYNIPQTDESTLMTIAYVMAAICALFMLPLCLMVCQWRCLRCLRQQHDDFADDISLLK DIP protein P56817 original sequence version: 161 377CE4C824ACEF05 377CE4C824ACEF05 itm2b_human Integral membrane protein 2B ITM2B BRI BRI2 Protein E25B Transmembrane protein BRI Immature BRI2 protein protein human Homo sapiens Human MVKVTFNSALAQKEAKKDEPKSGEEALIIPPDAVAVDCKDPDDVVPVGQRRAWCWCMCFGLAFMLAGVILGGAYLYKYFALQPDDVYYCGIKYIKDDVILNEPSADAPAALYQTIEENIKIFEEEEVEFISVPVPEFADSDPANIVHDFNKKLTAYLDLNLDKCYVIPLNTSIVMPPRNLLELLINIKAGTYLPQSYLIHEHMVITDRIENIDHLGFFIYRLCHDKETYKLQRRETIKGIQKREASNCFAIRHFENKFAVETLICS 3A7D8CA259F1F627 bace1-itm2b-1 3 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 2A To explore this possibility, we expressed in HEK293 cells mycBRI2 and BACE1 and performed immunoprecipitation of mycBRI2 from the cell extracts, using the antibody 9B11 against the myc epitope of BRI2 (Fig. 2A, panel a, lane 2). bace1-itm2b-2 2 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 2B In the reverse independent experiment (Fig. 2B) immunoprecipitation of BACE1 (panel a, lane 1) with the antibody 7520 resulted in co-precipitation of mycBRI2 (panel b, lane 1). bace1-itm2b-3 1 5 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level 6 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig. 2C To investigate whether the endogenous proteins interact, we immunoprecipitated endogenous BACE1 (Fig. 2C, panel a, lane 2) from extracts of the human neuroblastoma cell line SH-SY5Y, and showed that this resulted in co-precipitation of BRI2 (Fig. 2C, panel b, lane 2). itm2b-bace1-1 4 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed 6 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization Fig. 3 To further investigate the interaction between BRI2 and BACE1, we examined whether they co-localize in certain cellular compartments (Fig. 3).