Abstract
Wild type green fluorescent protein (GFP) from Aequorea victoria absorbs predominantly at 398 nm. Illumination with UV (254 nm) or visible (390 nm) light transforms this state (GFP398) into one absorbing at 483 nm (GFP483). Here we show that this photoconversion of GFP is a one-photon process that is paralleled by decarboxylation of Glu 222. We propose a mechanism in which decarboxylation is due to electron transfer between the γ-carboxylate of Glu 222 and the p-hydroxybenzylidene-imidazolidinone chromophore of GFP, followed by reverse transfer of an electron and a proton to the remaining carbon side chain atom of Glu 222. Oxidative decarboxylation of a γ-carboxylate represents a new type of posttranslational modification that may also occur in enzymes with high-potential reaction intermediates.
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Acknowledgements
We acknowledge E. Garman for helping establish the extent of X-ray induced structural damage and T. Davies for help with crystallographic computing. We thank A. Watts for help with ESR measurements, and C. Valance, R. Wilmouth and R. Aplin for access to their facilities. J.J.v.T. acknowledges financial support from the European Molecular Biology Organization and the Human Frontier Science Program Organization.
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van Thor, J., Gensch, T., Hellingwerf, K. et al. Phototransformation of green fluorescent protein with UV and visible light leads to decarboxylation of glutamate 222. Nat Struct Mol Biol 9, 37–41 (2002). https://doi.org/10.1038/nsb739
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DOI: https://doi.org/10.1038/nsb739
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