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Designing artificial fluorescent proteins and biosensors by genetically encoding molecular rotor-based amino acids
Nature Chemistry ( IF 19.2 ) Pub Date : 2024-11-28 , DOI: 10.1038/s41557-024-01675-x
Liming Hu, Wenbing Cao, Yihaofan Jiang, Wenkang Cai, Xiaoding Lou, Tao Liu

Fluorescent proteins are indispensable tools in biological and medical research. The fluorophores are typically encoded by the primary amino acid sequence, from which a fluorescent molecular rotor structure forms upon protein folding. Here, inspired by the fluorogenic property exhibited by native fluorophores, we designed a collection of fluorogenic non-canonical amino acids that feature this molecular rotor structure—termed fluorescent molecular rotor amino acids (FMR-AAs)—akin to native fluorescent protein fluorophores. By incorporating FMR-AAs into target proteins through an expanded genetic code, we use them as encoded fluorophore analogues within a confined protein microenvironment, thus readily transforming diverse non-fluorescent proteins into artificial fluorescent proteins. We also use FMR-AAs in selected proteins as sensitive fluorescent probes for monitoring protein–protein interactions and detecting protein conformational changes in vitro and in living cells. This approach enables the generation of artificial fluorescent proteins and the development of biosensors from potentially any protein of interest with minor modifications.



中文翻译:


通过对基于分子转子的氨基酸进行基因编码来设计人工荧光蛋白和生物传感器



荧光蛋白是生物学和医学研究中不可或缺的工具。荧光团通常由初级氨基酸序列编码,蛋白质折叠时从初级氨基酸序列形成荧光分子转子结构。在这里,受到天然荧光团所表现出的荧光特性的启发,我们设计了一系列荧光非经典氨基酸,这些氨基酸具有这种分子转子结构——称为荧光分子转子氨基酸 (FMR-AAs)——类似于天然荧光蛋白荧光基团。通过扩展的遗传密码将 FMR-AAs 掺入靶蛋白中,我们在受限的蛋白质微环境中将它们用作编码的荧光团类似物,从而很容易地将不同的非荧光蛋白转化为人工荧光蛋白。我们还使用选定蛋白质中的 FMR-AA 作为敏感的荧光探针,用于监测蛋白质-蛋白质相互作用并检测体外和活细胞中的蛋白质构象变化。这种方法能够产生人工荧光蛋白,并通过微小的修饰从任何感兴趣的蛋白质中开发生物传感器。

更新日期:2024-11-29
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