Proteolytically activated CRAC effectors through designed intramolecular inhibition

Jazbec, Vid (avtor), Jerala, Roman (avtor), Benčina, Mojca (avtor)

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.

• Jezik: Angleški jezik
• Ključne besede: STIM1, Orai, TEV protease, PPV protease, calcium signaling, coiled-coil peptides, bioluminescent probes, calcium, cell signaling, crystal cleavage, peptides, proteins
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: MF - Medicinska fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2022
• Št. strani: Str. 2756–2765
• Številčenje: Vol. 11, iss. 8
• PID: 20.500.12556/RUL-140820
• UDK: 577
• ISSN pri članku: 2161-5063
• DOI: 10.1021/acssynbio.2c00151
• COBISS.SI-ID: 115643651
• Datum objave v RUL: 19.09.2022

Gradivo je del revije

• Naslov: ACS synthetic biology
• Skrajšan naslov: ACS synth. biol.
• Založnik: American Chemical Society
• ISSN: 2161-5063
• COBISS.SI-ID: 519107609

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: biokemija, celice, kalcij, beljakovine, proteini