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Structural Basis of Photoswitching in Fluorescent Proteins

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Photoswitching Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1148))

Abstract

Fluorescent proteins have revolutionized life sciences because they allow noninvasive and highly specific labeling of biological samples. The subset of “phototransformable” fluorescent proteins recently attracted a widespread interest, as their fluorescence state can be modified upon excitation at defined wavelengths. The fluorescence emission of Reversibly Switchable Fluorescent Proteins (RSFPs), in particular, can be repeatedly switched on and off. RSFPs enable many new exciting modalities in fluorescence microscopy and biotechnology, including protein tracking, photochromic Förster Resonance Energy Transfer, super-resolution microscopy, optogenetics, and ultra-high-density optical data storage. Photoswitching in RSFPs typically results from chromophore cis–trans isomerization accompanied by a protonation change, but other switching schemes based on, e.g., chromophore hydration/dehydration have also been discovered. In this chapter, we review the main structural features at the basis of photoswitching in RSFPs.

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Abbreviations

GFP:

Green fluorescent protein

YFP:

Yellow fluorescent protein

FPs:

Fluorescent proteins

PTFPs:

Phototransformable fluorescent proteins

RSFPs:

Reversibly switchable fluorescent proteins

PCFPs:

Photoconvertible fluorescent proteins

PAFPs:

Photoactivatable fluorescent proteins

ESPT:

Excited state proton transfer

KIE:

Kinetic isotope effect

p-HBI:

4-(p-Hydroxybenzylidene)-5-imidazolinone

pcFRET:

Photochromic Förster resonance energy transfer

QM/MM:

Quantum mechanics/molecular mechanics

SMLM:

Single molecule localization microscopy

PALM:

Photoactivated localization microscopy

STORM:

Stochastic optical reconstruction microscopy

STED:

Stimulated emission depletion

RESOLFT:

Reversible saturable optical linear fluorescence transitions

SSIM:

Saturated structured illumination microscopy

(pc)SOFI:

(Photochromic) stochastic optical fluctuation imaging

IR:

Infra-red

HSQC:

Heteronuclear single quantum coherence

XFEL:

X-Ray-free electron laser

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Authors and Affiliations

  1. Institut de Biologie Structurale, Université Grenoble Alpes, Grenoble, France

    Chenxi Duan, Virgile Adam & Martin Byrdin

  2. Institut de Biologie Structurale (IBS),Université Grenoble Alpes, 41 Rue Jules Horowitz, F-38027, Grenoble, France

    Dominique Bourgeois

Authors
  1. Chenxi Duan
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  2. Virgile Adam
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  3. Martin Byrdin
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  4. Dominique Bourgeois
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Corresponding author

Correspondence to Dominique Bourgeois .

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Editors and Affiliations

  1. Institute of Anatomy and Cell Biology, University of Heidelberg, Heidelberg, Germany

    Sidney Cambridge

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Duan, C., Adam, V., Byrdin, M., Bourgeois, D. (2014). Structural Basis of Photoswitching in Fluorescent Proteins. In: Cambridge, S. (eds) Photoswitching Proteins. Methods in Molecular Biology, vol 1148. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0470-9_12

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  • DOI: https://doi.org/10.1007/978-1-4939-0470-9_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0469-3

  • Online ISBN: 978-1-4939-0470-9

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