纳米快速检测法：基于结构设计的仅含98个氨基酸的小型荧光团激活蛋白
NanoFAST: structure-based design of a small fluorogen-activating protein with only 98 amino acids.
作者信息
Mineev Konstantin S, Goncharuk Sergey A, Goncharuk Marina V, Povarova Natalia V, Sokolov Anatolii I, Baleeva Nadezhda S, Smirnov Alexander Yu, Myasnyanko Ivan N, Ruchkin Dmitry A, Bukhdruker Sergey, Remeeva Alina, Mishin Alexey, Borshchevskiy Valentin, Gordeliy Valentin, Arseniev Alexander S, Gorbachev Dmitriy A, Gavrikov Alexey S, Mishin Alexander S, Baranov Mikhail S
机构信息
Institute of Bioorganic Chemistry, Russian Academy of Sciences Miklukho-Maklaya 16/10 117997 Moscow Russia
Moscow Institute of Physics and Technology Dolgoprudny 141701 Russia.
出版信息
Chem Sci. 2021 Apr 8;12(19):6719-6725. doi: 10.1039/d1sc01454d.
One of the essential characteristics of any tag used in bioscience and medical applications is its size. The larger the label, the more it may affect the studied object, and the more it may distort its behavior. In this paper, using NMR spectroscopy and X-ray crystallography, we have studied the structure of fluorogen-activating protein FAST both in the apo form and in complex with the fluorogen. We showed that significant change in the protein occurs upon interaction with the ligand. While the protein is completely ordered in the complex, its apo form is characterized by higher mobility and disordering of its N-terminus. We used structural information to design the shortened FAST (which we named nanoFAST) by truncating 26 N-terminal residues. Thus, we created the shortest genetically encoded tag among all known fluorescent and fluorogen-activating proteins, which is composed of only 98 amino acids.
生物科学和医学应用中使用的任何标签的一个基本特征是其大小。标签越大，对研究对象的影响可能就越大，其行为的扭曲程度也可能越高。在本文中，我们利用核磁共振光谱和X射线晶体学研究了荧光激活蛋白FAST的无配体形式及其与荧光团复合物的结构。我们发现，蛋白质与配体相互作用时会发生显著变化。虽然该蛋白质在复合物中是完全有序的，但其无配体形式的特征是N端具有更高的流动性和无序性。我们利用结构信息，通过截短26个N端残基设计了缩短版的FAST（我们将其命名为nanoFAST）。因此，我们创造了所有已知荧光和荧光激活蛋白中最短的基因编码标签，它仅由98个氨基酸组成。
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