5WWK

Highly stable green fluorescent protein

5WWK の概要

• エントリーDOI: 10.2210/pdb5wwk/pdb
• 分子名称: Green fluorescent protein (2 entities in total)
• 機能のキーワード: chromophore modification, thermal stability fluorescence protein sensor, fluorescent protein
• 由来する生物種: Aequorea victoria
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 155263.13

構造登録者

Sriram, R.,George, A.,Kesavan, M.,Jaimohan, S.M.,Kamini, N.R.,Easwaramoorthi, S.,Ganesh, S.,Gunasekaran, K.,Ayyadurai, N. (登録日: 2017-01-02, 公開日: 2017-12-13, 最終更新日: 2024-10-30)

主引用文献

Augustine, G.,Raghavan, S.,NumbiRamudu, K.,Easwaramoorthi, S.,Shanmugam, G.,Seetharani Murugaiyan, J.,Gunasekaran, K.,Govind, C.,Karunakaran, V.,Ayyadurai, N.
Excited State Electronic Interconversion and Structural Transformation of Engineered Red-Emitting Green Fluorescent Protein Mutant.
J.Phys.Chem.B, 123:2316-2324, 2019

PubMed Abstract: Red fluorescent proteins with a large Stokes shift offer a limited autofluorescence background and are used in deep tissue imaging. Here, by introducing the free amino group in Aequorea victoria, the electrostatic charges of the p-hydroxybenzylidene imidazolinone chromophore of green fluorescent protein (GFP) have been altered resulting in an unusual, 85 nm red-shifted fluorescence. The structural and biophysical analysis suggested that the red shift is due to positional shift occupancy of Glu222 and Arg96, resulting in extended conjugation and a relaxed chromophore. Femtosecond transient absorption spectra exhibited that the excited state relaxation dynamics of red-shifted GFP (rGFP) (τ = 234 ps) are faster compared to the A. victoria green fluorescent protein (τ = 3.0 ns). The nanosecond time-resolved emission spectra of rGFP reveal the continuous spectral shift during emission by a solvent reorientation in the chromophore. Finally, the molecular dynamics simulations revealed the rearrangement of the hydrogen bond interactions in the chromophore vicinity, reshaping the symmetric distribution of van der Waals space to fine tune the GFP structure resulting from highly red-shifted rGFP.

PubMed: 30789731
DOI: 10.1021/acs.jpcb.8b10516

実験手法

X-RAY DIFFRACTION (3.199 Å)