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Photoconversion of the Green Fluorescent Protein and Related Proteins

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Fluorescent Proteins I

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 11))

Abstract

This review focuses on the mechanistic details of photochromic reactions of the green fluorescent protein (GFP) and also of its mutant derivatives and related fluorescent proteins. A number of distinct photochromic processes have so far been identified that have entirely different photochemical and chemical basis, which will be reviewed. In addition to bright fluorescence, the GFP from the jellyfish Aequorea victoria undergoes photochromic transformation with blue or UV illumination. The associated change in electronic absorption provides a spectroscopic contrast that can be used in fluorescence microscopy application to tag and track the movement of populations that are photoconverted. Key to the successful use of photoconversion for such microscopy experiments is in fact the relatively low quantum yield of the irreversible process. In the wild-type GFP, photoconversion is triggered by light-induced electron transfer from the buried anionic carboxylate of Glu222 to the optically excited protonated chromophore. An unstable carboxylate radical subsequently cleaves off a CO2 molecule in a “Kolbe” type reaction that has been trapped in a partially oriented site near the chromophore-binding site at 100K, as observed by low-temperature X-ray crystallography and cryo-infrared crystallography. Structural intermediates in the subsequent relaxation pathway involve motion of CO2, amino acids and H-bonded waters both in the chromophore vicinity and at longer range. This review provides an overview of the molecular characterisation using structural and spectroscopy methods of this photoconversion reaction of GFP. In addition, the mechanisms of photochromic reactions of mutants of GFP and related fluorescent proteins will be summarised and discussed. These include the cis–trans isomerisation and protonation changes in Dronpa, asFP595 and IrisFP and related proteins, light-induced maturation in aceGFPL, and photoinduced beta-elimination and backbone cleavage that leads to “green-to-red” photoconversion in EosFP, Kaede, IrisFP and KikGR.

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Acknowledgements

Jasper van Thor is a Royal Society University Research Fellow. JvT acknowledges support from the European Research Council (Grant Agreement N° 208650) and EPSRC (Grant Ref EP/I003304/1).

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    Jasper J. van Thor

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    Gregor Jung

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van Thor, J.J. (2011). Photoconversion of the Green Fluorescent Protein and Related Proteins. In: Jung, G. (eds) Fluorescent Proteins I. Springer Series on Fluorescence, vol 11. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2011_20

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