Photoactivation of LOV domains with chemiluminescence.

Autor: Ji Y; Institute of Physiological Chemistry and Pathobiochemistry, University of Münster 48149 Münster Germany wegnerse@uni-muenster.de., Heidari A; Institute of Physiological Chemistry and Pathobiochemistry, University of Münster 48149 Münster Germany wegnerse@uni-muenster.de., Nzigou Mombo B; Institute of Physiological Chemistry and Pathobiochemistry, University of Münster 48149 Münster Germany wegnerse@uni-muenster.de., Wegner SV; Institute of Physiological Chemistry and Pathobiochemistry, University of Münster 48149 Münster Germany wegnerse@uni-muenster.de.
Jazyk: angličtina
Zdroj: Chemical science [Chem Sci] 2023 Dec 11; Vol. 15 (3), pp. 1027-1038. Date of Electronic Publication: 2023 Dec 11 (Print Publication: 2024).
DOI: 10.1039/d3sc04815b
Abstrakt: Optogenetics has opened new possibilities in the remote control of diverse cellular functions with high spatiotemporal precision using light. However, delivering light to optically non-transparent systems remains a challenge. Here, we describe the photoactivation of light-oxygen-voltage-sensing domains (LOV domains) with in situ generated light from a chemiluminescence reaction between luminol and H 2 O 2 . This activation is possible due to the spectral overlap between the blue chemiluminescence emission and the absorption bands of the flavin chromophore in LOV domains. All four LOV domain proteins with diverse backgrounds and structures (iLID, BcLOV4, nMagHigh/pMagHigh, and VVDHigh) were photoactivated by chemiluminescence as demonstrated using a bead aggregation assay. The photoactivation with chemiluminescence required a critical light-output below which the LOV domains reversed back to their dark state with protein characteristic kinetics. Furthermore, spatially confined chemiluminescence produced inside giant unilamellar vesicles (GUVs) was able to photoactivate proteins both on the membrane and in solution, leading to the recruitment of the corresponding proteins to the GUV membrane. Finally, we showed that reactive oxygen species produced by neutrophil like cells can be converted into sufficient chemiluminescence to recruit the photoswitchable protein BcLOV4-mCherry from solution to the cell membrane. The findings highlight the utility of chemiluminescence as an endogenous light source for optogenetic applications, offering new possibilities for studying cellular processes in optically non-transparent systems.
Competing Interests: There are no conflicts to declare.
(This journal is © The Royal Society of Chemistry.)
Databáze: MEDLINE
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