8ZUR

Crystal structure of the F99S/M153T/V163A/T203V/E222Q variant of GFP at pH 5.0

8ZUR の概要

• エントリーDOI: 10.2210/pdb8zur/pdb
• 分子名称: Green fluorescent protein, CHLORIDE ION (3 entities in total)
• 機能のキーワード: green fluorescent protein, 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: 25911.91

構造登録者

Takeda, R.,Takeda, K. (登録日: 2024-06-10, 公開日: 2024-11-06, 最終更新日: 2026-03-18)

主引用文献

Takeda, R.,Tsutsumi, E.,Okatsu, K.,Fukai, S.,Takeda, K.
Structural characterization of green fluorescent protein in the I-state.
Sci Rep, 14:22832-22832, 2024

PubMed Abstract: Green fluorescent protein (GFP) is widely utilized as a fluorescent tag in biochemical fields. Whereas the intermediate (I) state has been proposed in the photoreaction cycle in addition to the A and B states, until now the structure of I has only been estimated by computational studies. In this paper, we report the crystal structures of the I stabilizing variants of GFP at high resolutions where respective atoms can be observed separately. Comparison with the structures in the other states highlights the structural feature of the I state. The side chain of one of the substituted residues, Val203, adopts the gauche- conformation observed for Thr203 in the A state, which is different from the B state. On the other hand, His148 interacts with the chromophore by ordinary hydrogen bonding with a distance of 2.85 Å, while the weaker interaction by longer distances is observed in the A state. Therefore, it was indicated that it is possible to distinguish three states A, B and I by the two hydrogen bond distances Oγ-Thr203···Oη-chromophore and Nδ1-His148···Oη-chromophore. We discuss the characteristics of the I intermediate of wild-type GFP on the bases of the structure estimated from the variant structures by quantum chemical calculations.

PubMed: 39353998
DOI: 10.1038/s41598-024-73696-y

実験手法

X-RAY DIFFRACTION (1.2 Å)