OzTracs: Optical Osmolality Reporters Engineered from Mechanosensitive Ion Channels
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biomolecules-12-00787.pdf
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Author(s)
Kleist, Thomas J.
Lin, I Winnie
Xu, Sophia
Maksaev, Grigory
Sadoine, Mayuri
Haswell, Elizabeth S.
Frommer, Wolf B.
Wudick, Michael M.
Date Issued
June 4, 2022
Publisher
Multidisciplinary Digital Publishing Institute
Citation
Biomolecules 12 (6): 787 (2022)
Version
Final published version
Abstract
Interactions between physical forces and membrane proteins underpin many forms of environmental sensation and acclimation. Microbes survive osmotic stresses with the help of mechanically gated ion channels and osmolyte transporters. Plant mechanosensitive ion channels have been shown to function in defense signaling. Here, we engineered genetically encoded osmolality sensors (OzTracs) by fusing fluorescent protein spectral variants to the mechanosensitive ion channels MscL from E. coli or MSL10 from A. thaliana. When expressed in yeast cells, the OzTrac sensors reported osmolality changes as a proportional change in the emission ratio of the two fluorescent protein domains. Live-cell imaging revealed an accumulation of fluorescent sensors in internal aggregates, presumably derived from the endomembrane system. Thus, OzTrac sensors serve as osmolality-dependent reporters through an indirect mechanism, such as effects on molecular crowding or fluorophore solvation.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.3390/biom12060787
