5Y8R

ZsYellow at pH 3.5

5Y8R の概要

• エントリーDOI: 10.2210/pdb5y8r/pdb
• 分子名称: GFP-like fluorescent chromoprotein FP538 (2 entities in total)
• 機能のキーワード: zsyellow, ph, fluorescent protein
• 由来する生物種: Zoanthus sp. (Green polyp)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 26416.41

構造登録者

Bae, J.E.,Kim, I.J.,Nam, K.H. (登録日: 2017-08-21, 公開日: 2017-09-13, 最終更新日: 2026-03-18)

主引用文献

Bae, J.E.,Kim, I.J.,Nam, K.H.
Disruption of the hydrogen bonding network determines the pH-induced non-fluorescent state of the fluorescent protein ZsYellow by protonation of Glu221.
Biochem. Biophys. Res. Commun., 493:562-567, 2017

PubMed Abstract: Many fluorescent proteins (FPs) exhibit fluorescence quenching at a low pH. This pH-induced non-fluorescent state of an FP serves as a useful indicator of the cellular pH. ZsYellow is widely used as an optical marker in molecular biology, but its pH-induced non-fluorescent state has not been characterized. Here, we report the pH-dependent spectral properties of ZsYellow, which exhibited the pH-induced non-fluorescence state at a pH below 4.0. We determined the crystal structures of ZsYellow at pH 3.5 (non-fluorescence state) and 8.0 (fluorescence state), which revealed the cis-configuration of the chromophore without pH-induced isomerization. In the non-fluorescence state, Arg95, which is involved in stabilization of the exited state of the chromophore, was found to more loosely interact with the carbonyl oxygen atom of the chromophore when compared to the interaction at pH 8.0. In the fluorescence state, Glu221, which is involved in the hydrogen bonding network around the chromophore, stably interacted with Gln42 and His202. By contrast, in the non-fluorescence state, the protonated conserved Glu221 residue exhibited a large conformational change and was separated from His202 by 5.46 Å, resulting in breakdown of the hydrogen bond network. Our results provide insight into the critical role of the conserved Glu221 residue for generating the pH-induced non-fluorescent state.

PubMed: 28867188
DOI: 10.1016/j.bbrc.2017.08.152

実験手法

X-RAY DIFFRACTION (2.3 Å)