5O8C

Composite structure of rsEGFP2 1ps following 400nm-laser irradiation of the off-state.

5O8C の概要

• エントリーDOI: 10.2210/pdb5o8c/pdb
• 関連するPDBエントリー: 5DTX 5DTY
• 分子名称: Green fluorescent protein (2 entities in total)
• 機能のキーワード: photoswitching on state fluorescent state chromophore sfx serial femtosecond crystallography cxi lcls time resolved crystallography chromohore isomerisation, fluorescent protein
• 由来する生物種: Aequorea victoria (Jellyfish)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 28099.69

構造登録者

Coquelle, N.,Sliwa, M.,Woodhouse, J.,Schiro, G.,Adam, V.,Aquila, A.,Barends, T.R.M.,Boutet, S.,Byrdin, M.,Carbajo, S.,De la Mora, E.,Doak, R.B.,Feliks, M.,Fieschi, F.,Foucar, L.,Guillon, V.,Hilpert, M.,Hunter, M.,Jakobs, S.,Koglin, J.E.,Kovacsova, G.,Lane, T.J.,Levy, B.,Liang, M.,Nass, K.,Ridard, J.,Robinson, J.S.,Roome, C.M.,Ruckebusch, C.,Seaberg, M.,Thepaut, M.,Cammarata, M.,Demachy, I.,Field, M.,Shoeman, R.L.,Bourgeois, D.,Colletier, J.P.,Schlichting, I.,Weik, M. (登録日: 2017-06-12, 公開日: 2017-12-06, 最終更新日: 2024-01-17)

主引用文献

Coquelle, N.,Sliwa, M.,Woodhouse, J.,Schiro, G.,Adam, V.,Aquila, A.,Barends, T.R.M.,Boutet, S.,Byrdin, M.,Carbajo, S.,De la Mora, E.,Doak, R.B.,Feliks, M.,Fieschi, F.,Foucar, L.,Guillon, V.,Hilpert, M.,Hunter, M.S.,Jakobs, S.,Koglin, J.E.,Kovacsova, G.,Lane, T.J.,Levy, B.,Liang, M.,Nass, K.,Ridard, J.,Robinson, J.S.,Roome, C.M.,Ruckebusch, C.,Seaberg, M.,Thepaut, M.,Cammarata, M.,Demachy, I.,Field, M.,Shoeman, R.L.,Bourgeois, D.,Colletier, J.P.,Schlichting, I.,Weik, M.
Chromophore twisting in the excited state of a photoswitchable fluorescent protein captured by time-resolved serial femtosecond crystallography.
Nat Chem, 10:31-37, 2018

PubMed Abstract: Chromophores absorb light in photosensitive proteins and thereby initiate fundamental biological processes such as photosynthesis, vision and biofluorescence. An important goal in their understanding is the provision of detailed structural descriptions of the ultrafast photochemical events that they undergo, in particular of the excited states that connect chemistry to biological function. Here we report on the structures of two excited states in the reversibly photoswitchable fluorescent protein rsEGFP2. We populated the states through femtosecond illumination of rsEGFP2 in its non-fluorescent off state and observed their build-up (within less than one picosecond) and decay (on the several picosecond timescale). Using an X-ray free-electron laser, we performed picosecond time-resolved crystallography and show that the hydroxybenzylidene imidazolinone chromophore in one of the excited states assumes a near-canonical twisted configuration halfway between the trans and cis isomers. This is in line with excited-state quantum mechanics/molecular mechanics and classical molecular dynamics simulations. Our new understanding of the structure around the twisted chromophore enabled the design of a mutant that displays a twofold increase in its off-to-on photoswitching quantum yield.

PubMed: 29256511
DOI: 10.1038/nchem.2853

実験手法

X-RAY DIFFRACTION (1.7 Å)