In Vivo Plasmonic Enhancement of the Photocycle Dynamics and Fluorescence of Genetically Encoded Voltage Indicators

Locarno, M.; Dong, Q.; Meng, X.; Glessi, C.; Hettema, N.; Brandsma, N.; Blokhuizen, S.; Castaneda, A.; Schmidt, T.; Ganapathy, S.; Post, M.; van Roemburg, L.; Xu, B.; Cho, C.-T.; Laan, L.; Chien, M.-P.; Brinks, D.

doi:10.1101/2024.02.07.579195

bioRxiv · 10.1101/2024.02.07.579195

In Vivo Plasmonic Enhancement of the Photocycle Dynamics and Fluorescence of Genetically Encoded Voltage Indicators

Locarno, M.·Dong, Q.·Meng, X.·Glessi, C.·Hettema, N.·Brandsma, N.·Blokhuizen, S.·Castaneda, A.·Schmidt, T.·Ganapathy, S.·Post, M.·van Roemburg, L.·Xu, B.·Cho, C.-T.·Laan, L.·Chien, M.-P.·Brinks, D.

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Abstract

Plasmonic nanoparticles are key components in nanophotonics1-11 and have applications in molecular detection and diagnostic platforms. 12-14 Coupling of dipoles to plasmonic antennas has allowed engineering of qualitatively altered behavior in isolated quantum systems, 8,15 but this promise has not been fulfilled in living systems, where the use of plasmonics is limited to tissue level applications of plasmonic particles as contrast agents16 or heat sources.17 Here we show that coupling to designed plasmonic nanoparticles can control the electrophysiological function of proteins in living cells. We designed nanostar geometries and achieved robust near-field coupling of these optimized nanoparticles to plasma membrane-localized Archaerhodopsin proteins. We enhanced the fluorescence of the coupled rhodopsins and increased their response speed to membrane voltage. We incorporated this plasmonic enhancement into a Markov chain photocycle model of the Archaerhodopsin mutant QuasAr6a, showing an increased fluorescence emission rate and manipulation of the protein dynamics through a combination of photocycle transition rate enhancements. These results represent the first scalable near-field coupling between plasmonic particles and fluorescent proteins in living cells. They show enhancement of protein function beyond what has been achievable through genetic engineering. This opens up a range of possibilities for engineering biological functionality through plasmonics.

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Locarno, M., Dong, Q., Meng, X., Glessi, C., Hettema, N., Brandsma, N., Blokhuizen, S., Castaneda, A., Schmidt, T., Ganapathy, S., Post, M., van Roemburg, L., Xu, B., Cho, C.-T., Laan, L., Chien, M.-P., Brinks, D.. 2024-02-09. In Vivo Plasmonic Enhancement of the Photocycle Dynamics and Fluorescence of Genetically Encoded Voltage Indicators. https://doi.org/10.1101/2024.02.07.579195

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