源自 mNeonGreen 的青色荧光蛋白。

Landon Zarowny, Damien Clavel, Ryan Johannson, Kévin Duarte, Hadrien Depernet, Jérôme Dupuy, Heather Baker, Alex Brown, Antoine Royant, Robert E Campbell
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引用次数: 0

摘要

mNeonGreen 是一种源自长尾鳕的工程化绿色荧光蛋白(GFP),是目前报道的维多利亚水母 GFP(avGFP)最明亮的荧光同源物之一。在这项工作中,我们研究了 mNeonGreen 的同源物在改变了发色团结构和荧光色调后是否还能保持这种明亮的荧光。我们发现,通过用其他芳香族氨基酸取代形成发色团的关键酪氨酸残基,mNeonGreen 对发色团修饰的耐受性普遍低于 avGFP。不过,我们最终还是成功地创造出了一种变体,命名为 NeonCyan1,它具有源自色氨酸的青色荧光蛋白(CFP)型发色团,以及另外两种具有不同光谱色调的突变体。通过对 NeonCyan1 及其变体进行结构、计算和光物理表征,我们深入了解了控制荧光发射颜色的因素。虽然我们不建议用 NeonCyan1 变体来替代现有的 CFP 变体,但我们证明了 NeonCyan1 变体有可能适用于活细胞成像应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Cyan fluorescent proteins derived from mNeonGreen.

mNeonGreen, an engineered green fluorescent protein (GFP) derived from lancelet, is one of the most brightly fluorescent homologs of Aequorea victoria jellyfish GFP (avGFP) yet reported. In this work, we investigated whether this bright fluorescence might be retained in homologs of mNeonGreen with modified chromophore structures and altered fluorescent hues. We found mNeonGreen to be generally less tolerant than avGFP to chromophore modification by substitution of the key chromophore-forming tyrosine residue with other aromatic amino acids. However, we were ultimately successful in creating a variant, designated as NeonCyan1, with a tryptophan-derived cyan fluorescent protein (CFP)-type chromophore, and two additional mutants with distinct spectral hues. Structural, computational, and photophysical characterization of NeonCyan1 and its variants provided insight into the factors that control the fluorescence emission color. Though not recommended as replacements for contemporary CFP variants, we demonstrate that NeonCyan1 variants are potentially suitable for live cell imaging applications.

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