Ultrasensitive measurements of microbial rhodopsin photocycles using photochromic FRET

Halil Bayraktar; Alexander P. Fields; Joel M. Kralj; John L. Spudich; Kenneth J. Rothschild; Adam E. Cohen

doi:10.1111/j.1751-1097.2011.01011.x

Ultrasensitive measurements of microbial rhodopsin photocycles using photochromic FRET

Halil Bayraktar, Alexander P. Fields, Joel M. Kralj, John L. Spudich, Kenneth J. Rothschild, Adam E. Cohen^*

^*Corresponding author for this work

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

28 Citations (Scopus)

Abstract

Microbial rhodopsins are an important class of light-activated transmembrane proteins whose function is typically studied on bulk samples. Herein, we apply photochromic fluorescence resonance energy transfer to investigate the dynamics of these proteins with sensitivity approaching the single-molecule limit. The brightness of a covalently linked organic fluorophore is modulated by changes in the absorption spectrum of the endogenous retinal chromophore that occur as the molecule undergoes a light-activated photocycle. We studied the photocycles of blue-absorbing proteorhodopsin and sensory rhodopsin II (SRII). Clusters of 2-3 molecules of SRII clearly showed a light-induced photocycle. Single molecules of SRII showed a photocycle upon signal averaging over several illumination cycles.

Original language	English
Pages (from-to)	90-97
Number of pages	8
Journal	Photochemistry and Photobiology
Volume	88
Issue number	1
DOIs	https://doi.org/10.1111/j.1751-1097.2011.01011.x
Publication status	Published - Jan 2012
Externally published	Yes

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title = "Ultrasensitive measurements of microbial rhodopsin photocycles using photochromic FRET",

abstract = "Microbial rhodopsins are an important class of light-activated transmembrane proteins whose function is typically studied on bulk samples. Herein, we apply photochromic fluorescence resonance energy transfer to investigate the dynamics of these proteins with sensitivity approaching the single-molecule limit. The brightness of a covalently linked organic fluorophore is modulated by changes in the absorption spectrum of the endogenous retinal chromophore that occur as the molecule undergoes a light-activated photocycle. We studied the photocycles of blue-absorbing proteorhodopsin and sensory rhodopsin II (SRII). Clusters of 2-3 molecules of SRII clearly showed a light-induced photocycle. Single molecules of SRII showed a photocycle upon signal averaging over several illumination cycles.",

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AU - Fields, Alexander P.

AU - Kralj, Joel M.

AU - Spudich, John L.

AU - Rothschild, Kenneth J.

AU - Cohen, Adam E.

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AB - Microbial rhodopsins are an important class of light-activated transmembrane proteins whose function is typically studied on bulk samples. Herein, we apply photochromic fluorescence resonance energy transfer to investigate the dynamics of these proteins with sensitivity approaching the single-molecule limit. The brightness of a covalently linked organic fluorophore is modulated by changes in the absorption spectrum of the endogenous retinal chromophore that occur as the molecule undergoes a light-activated photocycle. We studied the photocycles of blue-absorbing proteorhodopsin and sensory rhodopsin II (SRII). Clusters of 2-3 molecules of SRII clearly showed a light-induced photocycle. Single molecules of SRII showed a photocycle upon signal averaging over several illumination cycles.

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Ultrasensitive measurements of microbial rhodopsin photocycles using photochromic FRET

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