2VZX

Structural and spectroscopic characterization of photoconverting fluorescent protein Dendra2

2VZX の概要

• エントリーDOI: 10.2210/pdb2vzx/pdb
• 分子名称: Green fluorescent protein, TETRAETHYLENE GLYCOL, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: fluorescent protein, photoactivation, gfp-like protein, dendra2
• 由来する生物種: Dendronephthya sp. SSAL-2002
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 212453.00

構造登録者

Adam, V.,Nienhaus, K.,Bourgeois, D.,Nienhaus, G.U. (登録日: 2008-08-06, 公開日: 2009-06-09, 最終更新日: 2023-12-13)

主引用文献

Adam, V.,Nienhaus, K.,Bourgeois, D.,Nienhaus, G.U.
Structural Basis of Enhanced Photoconversion Yield in Green Fluorescent Protein-Like Protein Dendra2.
Biochemistry, 48:4905-, 2009

PubMed Abstract: Dendra2 is an engineered, monomeric GFP-like protein that belongs to a subclass of fluorescent proteins undergoing irreversible photoconversion from a green- to a red-emitting state upon exposure to purple-blue light. This photoinduced process occurs only in the neutral state of the chromophore and is known to result from backbone cleavage accompanied by an extension of the delocalized pi-electron system. We have measured the X-ray structure of the green species of Dendra2 and performed a comprehensive characterization of the optical absorption and fluorescence properties of the protein in both its green and red forms. The structure, which is very similar to those reported for the closely related proteins EosFP and Kaede, revealed a local structural change involving mainly Arg66 and a water molecule W4, which are part of a charged and hydrogen-bonded cluster of amino acids and water molecules next to the chromophore. Unlike in EosFP and Kaede, Arg66 of Dendra2 does not contribute to negative charge stabilization on the imidazolinone ring by hydrogen bonding to the imidazolinone carbonyl. This structural change may explain the blue shift of the absorption and emission bands, as well as the markedly higher pKs of the hydroxyphenyl moiety of the chromophore, which were determined as 7.1 and 7.5 for the green and red species, respectively. The action spectrum of photoconversion coincides with the absorption band of the neutral species. Consequently, its 20-fold enhancement in Dendra2 at physiological pH accounts for the higher photoconversion yield of this protein as compared to EosFP.

PubMed: 19371086
DOI: 10.1021/BI900383A

実験手法

X-RAY DIFFRACTION (2 Å)