6SM0

Venus 66 p-Azido-L-Phenylalanin (azF) variant, dark grown

6SM0 の概要

• エントリーDOI: 10.2210/pdb6sm0/pdb
• 分子名称: Green fluorescent protein, OXYGEN MOLECULE, ZINC ION, ... (5 entities in total)
• 機能のキーワード: venus 66 fluorescent protein, azf variant, aequoria victoria, gfp, non-canonical amino acid, 3d x-ray structure determination, 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: 25887.34

構造登録者

Rizkallah, P.J.,Al Maslookhi, H.S.,Jones, D.D. (登録日: 2019-08-21, 公開日: 2021-02-17, 最終更新日: 2024-01-24)

主引用文献

Auhim, H.S.,Grigorenko, B.L.,Harris, T.K.,Aksakal, O.E.,Polyakov, I.V.,Berry, C.,Gomes, G.D.P.,Alabugin, I.V.,Rizkallah, P.J.,Nemukhin, A.V.,Jones, D.D.
Stalling chromophore synthesis of the fluorescent protein Venus reveals the molecular basis of the final oxidation step.
Chem Sci, 12:7735-7745, 2021

PubMed Abstract: Fluorescent proteins (FPs) have revolutionised the life sciences, but the mechanism of chromophore maturation is still not fully understood. Here we show that incorporation of a photo-responsive non-canonical amino acid within the chromophore stalls maturation of Venus, a yellow FP, at an intermediate stage; a crystal structure indicates the presence of O located above a dehydrated enolate form of the imidazolone ring, close to the strictly conserved Gly67 that occupies a twisted conformation. His148 adopts an "open" conformation so forming a channel that allows O access to the immature chromophore. Absorbance spectroscopy supported by QM/MM simulations suggests that the first oxidation step involves formation of a hydroperoxyl intermediate in conjunction with dehydrogenation of the methylene bridge. A fully conjugated mature chromophore is formed through release of HO, both and . The possibility of interrupting and photochemically restarting chromophore maturation and the mechanistic insights open up new approaches for engineering optically controlled fluorescent proteins.

PubMed: 34168826
DOI: 10.1039/d0sc06693a

実験手法

X-RAY DIFFRACTION (1.91 Å)