Structural analysis of the bright monomeric yellow-green fluorescent protein mNeonGreen obtained by directed evolution.

Abstract : Until recently, genes coding for homologues of the autofluorescent protein GFP had only been identified in marine organisms from the phyla Cnidaria and Arthropoda. New fluorescent-protein genes have now been found in the phylum Chordata, coding for particularly bright oligomeric fluorescent proteins such as the tetrameric yellow fluorescent protein lanYFP from Branchiostoma lanceolatum. A successful monomerization attempt led to the development of the bright yellow-green fluorescent protein mNeonGreen. The structures of lanYFP and mNeonGreen have been determined and compared in order to rationalize the directed evolution process leading from a bright, tetrameric to a still bright, monomeric fluorescent protein. An unusual discolouration of crystals of mNeonGreen was observed after X-ray data collection, which was investigated using a combination of X-ray crystallography and UV-visible absorption and Raman spectroscopies, revealing the effects of specific radiation damage in the chromophore cavity. It is shown that X-rays rapidly lead to the protonation of the phenolate O atom of the chromophore and to the loss of its planarity at the methylene bridge.
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Acta crystallographica. Section D, Structural biology, International Union of Crystallography, 2016, 72 (12), pp.1298-1307. 〈10.1107/S2059798316018623〉
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Soumis le : mardi 17 janvier 2017 - 09:21:19
Dernière modification le : lundi 19 février 2018 - 14:34:03

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Damien Clavel, Guillaume Gotthard, David Von Stetten, Daniele De Sanctis, Hélène Pasquier, et al.. Structural analysis of the bright monomeric yellow-green fluorescent protein mNeonGreen obtained by directed evolution.. Acta crystallographica. Section D, Structural biology, International Union of Crystallography, 2016, 72 (12), pp.1298-1307. 〈10.1107/S2059798316018623〉. 〈hal-01437105〉

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