4B30

Structure of the mutant V44A of the fluorescent protein KillerRed

4B30 の概要

• エントリーDOI: 10.2210/pdb4b30/pdb
• 関連するPDBエントリー: 2WIQ 2WIS
• 分子名称: KILLERRED, SULFATE ION, CHLORIDE ION, ... (7 entities in total)
• 機能のキーワード: fluorescent protein, photobleaching, photoconversion, ros
• 由来する生物種: ANTHOMEDUSAE SP. DC-2005; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 53797.56

構造登録者

Carpentier, P.,de Rosny, E. (登録日: 2012-07-19, 公開日: 2013-01-16, 最終更新日: 2025-10-01)

主引用文献

De Rosny, E.,Carpentier, P.
Gfp-Like Phototransformation Mechanisms in the Cytotoxic Fluorescent Protein Killerred Unraveled by Structural and Spectroscopic Investigations.
J.Am.Chem.Soc., 134:18015-, 2012

PubMed Abstract: KillerRed (KR) is a red fluorescent protein recognized as an efficient genetically encoded photosensitizer. KR generates reactive oxygen species via a complex process of photoreactions, ending up in photobleaching, the mechanism of which remains obscure. In order to clarify these mechanisms, we focus on a single mutant V44A (A44-KR) exhibiting the solely green component of KR. We report on the laser-induced structural transformations of A44-KR at cryogenic temperature, which we have investigated by combining UV-vis fluorescence/absorption spectroscopy with X-ray crystallography. Like the well-known GFP, A44-KR possesses a mixture of protonated (A) absorbing at 397 and deprotonated (B) absorbing at 515 nm chromophores, which are stressed by intense prolonged violet and blue laser sources. Both illuminations directly drive the B-chromophores toward a bleached trans isomerized form. A-type chromophores are sensitive only to violet illumination and are phototransformed either into a deprotonated green fluorescent form by decarboxylation of E218 or into a bleached form with a disordered p-hydroxybenzylidene. In crystallo spectroscopy at cryo-temperature allowed the identification and dissection of an exhaustive scheme of intermediates and end-products resulting from the phototransformation of A44-KR. This constitutes a framework for understanding the photochemistry of the photosensitizer KillerRed.

PubMed: 23025285
DOI: 10.1021/JA3073337

実験手法

X-RAY DIFFRACTION (2.1 Å)