7RRH

Crystal structure of fast switching R66M/M159T mutant of fluorescent protein Dronpa (Dronpa2)

7RRH の概要

• エントリーDOI: 10.2210/pdb7rrh/pdb
• 分子名称: Fluorescent protein Dronpa (2 entities in total)
• 機能のキーワード: dronpa2, fluorescent protein
• 由来する生物種: Echinophyllia sp. SC22
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 173613.48

構造登録者

Lin, C.-Y.,Romei, M.G.,Mathews, I.I.,Boxer, S.G. (登録日: 2021-08-09, 公開日: 2021-10-13, 最終更新日: 2024-11-13)

主引用文献

Lin, C.Y.,Romei, M.G.,Mathews, I.I.,Boxer, S.G.
Energetic Basis and Design of Enzyme Function Demonstrated Using GFP, an Excited-State Enzyme.
J.Am.Chem.Soc., 144:3968-3978, 2022

PubMed Abstract: The past decades have witnessed an explosion of protein designs with a remarkable range of scaffolds. It remains challenging, however, to design catalytic functions that are competitive with naturally occurring counterparts as well as biomimetic or nonbiological catalysts. Although directed evolution often offers efficient solutions, the fitness landscape remains opaque. Green fluorescent protein (GFP), which has revolutionized biological imaging and assays, is one of the most redesigned proteins. While not an enzyme in the conventional sense, GFPs feature competing excited-state decay pathways with the same steric and electrostatic origins as conventional ground-state catalysts, and they exert exquisite control over multiple reaction outcomes through the same principles. Thus, GFP is an "excited-state enzyme". Herein we show that rationally designed mutants and hybrids that contain environmental mutations and substituted chromophores provide the basis for a quantitative model and prediction that describes the influence of sterics and electrostatics on excited-state catalysis of GFPs. As both perturbations can selectively bias photoisomerization pathways, GFPs with fluorescence quantum yields (FQYs) and photoswitching characteristics tailored for specific applications could be predicted and then demonstrated. The underlying energetic landscape, readily accessible via spectroscopy for GFPs, offers an important missing link in the design of protein function that is generalizable to catalyst design.

PubMed: 35200017
DOI: 10.1021/jacs.1c12305

実験手法

X-RAY DIFFRACTION (1.747 Å)