Moerke Nathan J
Harvard Medical School, Boston, Massachusetts.
Curr Protoc Chem Biol. 2009 Dec 1;1(1):1-15. doi: 10.1002/9780470559277.ch090102.
The technique of fluorescence polarization (FP) is based on the observation that when a fluorescently labeled molecule is excited by polarized light, it emits light with a degree of polarization that is inversely proportional to the rate of molecular rotation. This property of fluorescence can be used to measure the interaction of a small labeled ligand with a larger protein and provides a basis for direct and competition binding assays. FP assays are readily adaptable to a high-throughput format, have been used successfully in screens directed against a wide range of targets, and are particularly valuable in screening for inhibitors of protein-protein and protein-nucleic acid interactions when a small binding epitope can be identified for one of the partners. The protocols in this article describe a general procedure for development of FP assays to monitor binding of such a peptide or oligonucleotide to a protein of interest. Curr. Protoc. Chem Biol. 1:1-15. © 2009 by John Wiley & Sons, Inc.
荧光偏振（FP）技术基于这样的观察结果：当一个荧光标记的分子被偏振光激发时，它发射出的光的偏振程度与分子旋转速率成反比。荧光的这一特性可用于测量小分子标记配体与大分子蛋白质之间的相互作用，并为直接和竞争结合测定提供了基础。FP测定很容易适应高通量形式，已成功用于针对多种靶标的筛选，并且在筛选蛋白质-蛋白质和蛋白质-核酸相互作用的抑制剂时特别有价值，前提是可以为其中一个相互作用伙伴鉴定出一个小的结合表位。本文中的方案描述了开发FP测定以监测此类肽或寡核苷酸与感兴趣蛋白质结合的一般程序。《化学生物学实验指南》1:1 - 15。©2009约翰威立国际出版公司。
