Structure of a fluorescent protein from Aequorea victoria bearing the obligate-monomer mutation A206K.

IF 0.9 4区 生物学
David von Stetten, Marjolaine Noirclerc-Savoye, Joachim Goedhart, Theodorus W J Gadella, Antoine Royant
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引用次数: 0

Abstract

The green fluorescent protein (GFP) from the jellyfish Aequoria victoria has been shown to dimerize at high concentrations, which could lead to artefacts in imaging experiments. To ensure a truly monomeric state, an A206K mutation has been introduced into most of its widely used variants, with minimal effect on the spectroscopic properties. Here, the first structure of one of these variants, the cyan fluorescent protein mTurquoise, is presented and compared with that of its dimeric version mTurquoise-K206A. No significant structural change is detected in the chromophore cavity, reinforcing the notion that this mutation is spectroscopically silent and validating that the structural analysis performed on dimeric mutants also applies to monomeric versions. Finally, it is explained why cyan versions of GFP containing the Y66W and N146I mutations do not require the A206K mutation to prevent dimerization at high concentrations.

Abstract Image

带有强制性单体突变 A206K 的 Aequorea victoria 荧光蛋白的结构。
来自维多利亚水母(Aequoria victoria)的绿色荧光蛋白(GFP)已被证明在高浓度下会产生二聚体,这可能会导致成像实验中的伪影。为了确保真正的单体状态,在大多数广泛使用的变体中都引入了 A206K 突变,但对光谱特性的影响很小。本文首次展示了这些变体之一--青色荧光蛋白 mTurquoise 的结构,并与其二聚体版本 mTurquoise-K206A 的结构进行了比较。在发色团空腔中没有检测到明显的结构变化,这进一步证实了这种突变在光谱学上是无声的,同时也验证了对二聚体突变体进行的结构分析同样适用于单体突变体。最后,还解释了为什么含有 Y66W 和 N146I 突变的青色 GFP 不需要 A206K 突变来防止高浓度下的二聚化。
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来源期刊
自引率
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0
审稿时长
2-4 weeks
期刊介绍: Acta Crystallographica Section F is a rapid structural biology communications journal. Articles on any aspect of structural biology, including structures determined using high-throughput methods or from iterative studies such as those used in the pharmaceutical industry, are welcomed by the journal. The journal offers the option of open access, and all communications benefit from unlimited free use of colour illustrations and no page charges. Authors are encouraged to submit multimedia content for publication with their articles. Acta Cryst. F has a dedicated online tool called publBio that is designed to make the preparation and submission of articles easier for authors.
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