Photocrosslinking-induced CRAC channel-like Orai1 activation independent of STIM1

Lena Maltan; Sarah Weiß; Hadil Najjar; Melanie Leopold; Sonja Lindinger; Carmen Höglinger; Lorenz Höbarth; Matthias Sallinger; Herwig Grabmayr; Sascha Berlansky; Denis Krivic; Valentina Hopl; Anna Blaimschein; Marc Fahrner; Irene Frischauf; Adela Tiffner; Isabella Derler

doi:10.1038/s41467-023-36458-4

Photocrosslinking-induced CRAC channel-like Orai1 activation independent of STIM1

Lena Maltan, Sarah Weiß, Hadil Najjar, Melanie Leopold, Sonja Lindinger, Carmen Höglinger, Lorenz Höbarth, Matthias Sallinger, Herwig Grabmayr, Sascha Berlansky, Denis Krivic, Valentina Hopl, Anna Blaimschein, Marc Fahrner, Irene Frischauf, Adela Tiffner, Isabella Derler

Department of Molecular and Membrane Biophysics

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Article number	1286
Pages (from-to)	1286
Number of pages	14
Journal	Nature Communications
Volume	14
Issue number	1
DOIs	https://doi.org/10.1038/s41467-023-36458-4
Publication status	Published - 08 Mar 2023

Fields of science

103 Physics, Astronomy
106006 Biophysics

JKU Focus areas

Sustainable Development: Responsible Technologies and Management

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10.1038/s41467-023-36458-4

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Maltan, L., Weiß, S., Najjar, H., Leopold, M., Lindinger, S., Höglinger, C., Höbarth, L., Sallinger, M., Grabmayr, H., Berlansky, S., Krivic, D., Hopl, V., Blaimschein, A., Fahrner, M., Frischauf, I., Tiffner, A., & Derler, I. (2023). Photocrosslinking-induced CRAC channel-like Orai1 activation independent of STIM1. Nature Communications, 14(1), 1286. Article 1286. https://doi.org/10.1038/s41467-023-36458-4

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title = "Photocrosslinking-induced CRAC channel-like Orai1 activation independent of STIM1",

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.",

author = "Lena Maltan and Sarah Wei{\ss} and Hadil Najjar and Melanie Leopold and Sonja Lindinger and Carmen H{\"o}glinger and Lorenz H{\"o}barth and Matthias Sallinger and Herwig Grabmayr and Sascha Berlansky and Denis Krivic and Valentina Hopl and Anna Blaimschein and Marc Fahrner and Irene Frischauf and Adela Tiffner and Isabella Derler",

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doi = "10.1038/s41467-023-36458-4",

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Maltan, L, Weiß, S, Najjar, H, Leopold, M, Lindinger, S, Höglinger, C, Höbarth, L, Sallinger, M, Grabmayr, H, Berlansky, S, Krivic, D, Hopl, V, Blaimschein, A, Fahrner, M , Frischauf, I , Tiffner, A & Derler, I 2023, 'Photocrosslinking-induced CRAC channel-like Orai1 activation independent of STIM1', Nature Communications, vol. 14, no. 1, 1286, pp. 1286. https://doi.org/10.1038/s41467-023-36458-4

TY - JOUR

T1 - Photocrosslinking-induced CRAC channel-like Orai1 activation independent of STIM1

AU - Maltan, Lena

AU - Weiß, Sarah

AU - Najjar, Hadil

AU - Leopold, Melanie

AU - Lindinger, Sonja

AU - Höglinger, Carmen

AU - Höbarth, Lorenz

AU - Sallinger, Matthias

AU - Grabmayr, Herwig

AU - Berlansky, Sascha

AU - Krivic, Denis

AU - Hopl, Valentina

AU - Blaimschein, Anna

AU - Fahrner, Marc

AU - Frischauf, Irene

AU - Tiffner, Adela

AU - Derler, Isabella

PY - 2023/3/8

Y1 - 2023/3/8

N2 - 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.

AB - 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.

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Photocrosslinking-induced CRAC channel-like Orai1 activation independent of STIM1

Abstract

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