Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, 3001 Leuven, Belgium.
Microbiology (Reading). 2024 Mar;170(3). doi: 10.1099/mic.0.001451.
Fluorescent proteins (FPs) have always been a crucial part of molecular research in life sciences, including the research into the human fungal pathogen but have obvious shortcomings such as their relatively large size and long maturation time. However, the next generation of FPs overcome these issues and rely on the binding of a fluorogen for the protein to become fluorescently active. This generation of FPs includes the improved version of Fluorescence activating and Absorption Shifting Tag (iFAST). The binding between the fluorogen and the iFAST protein is reversible, thus resulting in reversible fluorescence. The fluorogens of iFAST are analogues of 4-hydroxylbenzylidene-rhodanine (HBR). These HBR analogues differ in spectral properties depending on functional group substitutions, which gives the iFAST system flexibility in terms of absorbance and emission maxima. In this work we describe and illustrate the application of iFAST as a protein tag and its reversible multi-colour characteristics in .
荧光蛋白（FPs）一直是生命科学中分子研究的重要组成部分，包括对人类真菌病原体的研究，但它们存在明显的缺点，如相对较大的尺寸和较长的成熟时间。然而，下一代的 FPs 克服了这些问题，依赖于荧光团与蛋白质的结合使蛋白质具有荧光活性。这一代的 FPs 包括改良版的荧光激活和吸收移位标签（iFAST）。荧光团与 iFAST 蛋白之间的结合是可逆的，从而导致荧光的可逆性。iFAST 的荧光团是 4-羟基苯亚甲基罗丹宁（HBR）的类似物。这些 HBR 类似物在光谱性质上因官能团取代而有所不同，这使得 iFAST 系统在吸收和发射最大值方面具有灵活性。在这项工作中，我们描述并说明了 iFAST 作为一种蛋白质标签的应用及其在 中的可逆多色特性。
