20.500.12556/RUL-140820 Proteolytically activated CRAC effectors through designed intramolecular inhibition Highly regulated intracellular calcium entry affects numerous cellular physiological events. External regulation of intracellular calcium signaling presents a great opportunity for the artificial regulation of cellular activity. Calcium entry can be mediated by STIM proteins interacting with Orai calcium channels; therefore, the STIM1−Orai1 pair has become a tool for artificially modulating calcium entry. We report on an innovative genetically engineered protease-activated Orai activator called PACE. CAD self-dimerization and activation were inhibited with a coiled-coil forming peptide pair linked to CAD via a protease cleavage site. PACE generated sustained calcium entry after its activation with a reconstituted split protease. We also generated PACE, whose transcriptional activation of NFAT was triggered by PPV or TEV protease. Using PACE, we successfully activated the native NFAT signaling pathway and the production of cytokines in a T-cell line. PACE represents a useful tool for generating sustained calcium entry to initiate calcium-dependent protein translation. PACE provides a promising template for the construction of links between various protease activation pathways and calcium signaling. STIM1 Orai TEV protease PPV protease calcium signaling coiled-coil peptides bioluminescent probes calcium cell signaling crystal cleavage peptides proteins biokemija celice kalcij beljakovine proteini true false true Angleški jezik Slovenski jezik Članek v reviji 2022-09-19 08:26:12 2022-09-19 08:26:15 2022-09-24 04:12:17 0000-00-00 00:00:00 2022 0 0 Str. 2756–2765 iss. 8 Vol. 11 2022 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577 2161-5063 10.1021/acssynbio.2c00151 115643651 acssynbio.2c00151_Proteolytically_activated_CRAC_effectors_through_designed_intramolecular_inhibition.pdf acssynbio.2c00151_Proteolytically_activated_CRAC_effectors_through_designed_intramolecular_inhibition.pdf 1 55A325DB1F5769259A7876FEF0E3CF65 18561abea6adc3c3c8acc34d10a564b08f96cc3f40d04321eedcac9d16d6e797 8ea82fdd-3b28-11ed-9c94-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=162063 https://pubs.acs.org/doi/10.1021/acssynbio.2c00151 1 e2ef04af-37e3-11ed-92af-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=161616 Medicinska fakulteta 0 0 0