7UGR

Crystal structure of hyperfolder YFP

7UGR の概要

• エントリーDOI: 10.2210/pdb7ugr/pdb
• 分子名称: Hyperfolder yellow fluorescent protein, TETRAETHYLENE GLYCOL, DI(HYDROXYETHYL)ETHER, ... (5 entities in total)
• 機能のキーワード: hfyfp, hyperfolder, fluorescent protein, superfolder
• 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: 27467.98

構造登録者

Campbell, B.C.,Liu, C.F.,Petsko, G.A. (登録日: 2022-03-25, 公開日: 2022-10-26, 最終更新日: 2024-10-16)

主引用文献

Campbell, B.C.,Paez-Segala, M.G.,Looger, L.L.,Petsko, G.A.,Liu, C.F.
Chemically stable fluorescent proteins for advanced microscopy.
Nat.Methods, 19:1612-1621, 2022

PubMed Abstract: We report the rational engineering of a remarkably stable yellow fluorescent protein (YFP), 'hyperfolder YFP' (hfYFP), that withstands chaotropic conditions that denature most biological structures within seconds, including superfolder green fluorescent protein (GFP). hfYFP contains no cysteines, is chloride insensitive and tolerates aldehyde and osmium tetroxide fixation better than common fluorescent proteins, enabling its use in expansion and electron microscopies. We solved crystal structures of hfYFP (to 1.7-Å resolution), a monomeric variant, monomeric hyperfolder YFP (1.6 Å) and an mGreenLantern mutant (1.2 Å), and then rationally engineered highly stable 405-nm-excitable GFPs, large Stokes shift (LSS) monomeric GFP (LSSmGFP) and LSSA12 from these structures. Lastly, we directly exploited the chemical stability of hfYFP and LSSmGFP by devising a fluorescence-assisted protein purification strategy enabling all steps of denaturing affinity chromatography to be visualized using ultraviolet or blue light. hfYFP and LSSmGFP represent a new generation of robustly stable fluorescent proteins developed for advanced biotechnological applications.

PubMed: 36344833
DOI: 10.1038/s41592-022-01660-7

実験手法

X-RAY DIFFRACTION (1.74 Å)