Royer Catherine A, Scarlata Suzanne F
Centre de Biochimie Structurale, 29 rue de Navacelles, 34090 Montpellier Cedex, France.
Methods Enzymol. 2008;450:79-106. doi: 10.1016/S0076-6879(08)03405-8.
This review is conceived as an introductory text to aid in the understanding and conception of fluorescence-based measurements of biomolecular interactions. The major fluorescence observables are introduced briefly. Next, the criteria that are involved in the choice of the fluorescent probe are discussed in terms of their advantages and disadvantages for different types of experiments. The last sections deal with the experimental design for fluorescence-based assays aimed at detecting different types of biomolecular interactions. Included in our examples are protein-ligand interactions, protein-nucleic acid interactions, aqueous phase protein-protein interactions and protein interactions in or at the cell membrane. We hope that this introduction will be of use to students and researchers considering the use of fluorescence in their work.
本综述旨在作为一篇介绍性文章,以帮助理解和构思基于荧光的生物分子相互作用测量方法。简要介绍了主要的荧光观测指标。接下来,根据荧光探针在不同类型实验中的优缺点,讨论了选择荧光探针所涉及的标准。最后几部分涉及基于荧光分析的实验设计,旨在检测不同类型的生物分子相互作用。我们的示例包括蛋白质-配体相互作用、蛋白质-核酸相互作用、水相蛋白质-蛋白质相互作用以及细胞膜内或膜上的蛋白质相互作用。我们希望这篇引言对考虑在工作中使用荧光技术的学生和研究人员有所帮助。