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- Title
Photocrosslinking-induced CRAC channel-like Orai1 activation independent of STIM1.
- Authors
Maltan, Lena; Weiß, Sarah; Najjar, Hadil; Leopold, Melanie; Lindinger, Sonja; Höglinger, Carmen; Höbarth, Lorenz; Sallinger, Matthias; Grabmayr, Herwig; Berlansky, Sascha; Krivic, Denis; Hopl, Valentina; Blaimschein, Anna; Fahrner, Marc; Frischauf, Irene; Tiffner, Adéla; Derler, Isabella
- Abstract
Ca2+ release-activated Ca2+ (CRAC) channels, indispensable for the immune system and various other human body functions, consist of two transmembrane (TM) proteins, the Ca2+-sensor STIM1 in the ER membrane and the Ca2+ ion channel Orai1 in the plasma membrane. Here we employ genetic code expansion in mammalian cell lines to incorporate the photocrosslinking unnatural amino acids (UAA), p-benzoyl-L-phenylalanine (Bpa) and p-azido-L-phenylalanine (Azi), into the Orai1 TM domains at different sites. Characterization of the respective UAA-containing Orai1 mutants using Ca2+ imaging and electrophysiology reveal that exposure to UV light triggers a range of effects depending on the UAA and its site of incorporation. In particular, photoactivation at A137 using Bpa in Orai1 activates Ca2+ currents that best match the biophysical properties of CRAC channels and are capable of triggering downstream signaling pathways such as nuclear factor of activated T-cells (NFAT) translocation into the nucleus without the need for the physiological activator STIM1. The Ca2+ ion channel Orai1 is crucial in immune cells. Here, the authors applied genetic code expansion to transfer light-sensitivity to the Orai1 channel and achieved precise control over its function.
- Subjects
ION channels; CALCIUM channels; NUCLEAR factor of activated T-cells; GENETIC code; CELL membranes; PHOTOCROSSLINKING
- Publication
Nature Communications, 2023, Vol 14, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-023-36458-4