8AM4

Cl-rsEGFP2 Long Wavelength Structure

8AM4 の概要

• エントリーDOI: 10.2210/pdb8am4/pdb
• 分子名称: Green fluorescent protein (1 entity in total)
• 機能のキーワード: gfp-like protein, beta-barrel, bioluminescence, fluorescent protein
• 由来する生物種: Aequorea victoria
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 28566.65

構造登録者

Orr, C.M.,Fadini, A.,van Thor, J. (登録日: 2022-08-02, 公開日: 2023-08-02, 最終更新日: 2024-01-31)

主引用文献

Fadini, A.,Hutchison, C.D.M.,Morozov, D.,Chang, J.,Maghlaoui, K.,Perrett, S.,Luo, F.,Kho, J.C.X.,Romei, M.G.,Morgan, R.M.L.,Orr, C.M.,Cordon-Preciado, V.,Fujiwara, T.,Nuemket, N.,Tosha, T.,Tanaka, R.,Owada, S.,Tono, K.,Iwata, S.,Boxer, S.G.,Groenhof, G.,Nango, E.,van Thor, J.J.
Serial Femtosecond Crystallography Reveals that Photoactivation in a Fluorescent Protein Proceeds via the Hula Twist Mechanism.
J.Am.Chem.Soc., 2023

PubMed Abstract: Chromophore photoisomerization is a fundamental process in chemistry and in the activation of many photosensitive proteins. A major task is understanding the effect of the protein environment on the efficiency and direction of this reaction compared to what is observed in the gas and solution phases. In this study, we set out to visualize the hula twist (HT) mechanism in a fluorescent protein, which is hypothesized to be the preferred mechanism in a spatially constrained binding pocket. We use a chlorine substituent to break the twofold symmetry of the embedded phenolic group of the chromophore and unambiguously identify the HT primary photoproduct. Through serial femtosecond crystallography, we then track the photoreaction from femtoseconds to the microsecond regime. We observe signals for the photoisomerization of the chromophore as early as 300 fs, obtaining the first experimental structural evidence of the HT mechanism in a protein on its femtosecond-to-picosecond timescale. We are then able to follow how chromophore isomerization and twisting lead to secondary structure rearrangements of the protein β-barrel across the time window of our measurements.

PubMed: 37418747
DOI: 10.1021/jacs.3c02313

実験手法

X-RAY DIFFRACTION (2.02 Å)