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Structural Basis of Photoswitching in Fluorescent Proteins

  • Chenxi Duan
  • Virgile Adam
  • Martin Byrdin
  • Dominique BourgeoisEmail author
Part of the Methods in Molecular Biology book series (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 cistrans 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.

Key words

Fluorescent proteins Photoswitching Dronpa RSFPs Protein dynamics cistrans Isomerization Proton transfer Super-resolution microscopy 

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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Chenxi Duan
    • 1
  • Virgile Adam
    • 1
  • Martin Byrdin
    • 1
  • Dominique Bourgeois
    • 2
    Email author
  1. 1.Institut de Biologie Structurale, Université Grenoble AlpesGrenobleFrance
  2. 2.Institut de Biologie Structurale (IBS),Université Grenoble AlpesGrenobleFrance

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