Leifer Becky S, Doyle Shelby K, Richters André, Evans Helen L, Koehler Angela N
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United States; Center for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, MA, United States; The Broad Institute of MIT and Harvard, Cambridge, MA, United States.
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, United States; Center for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, MA, United States; The Broad Institute of MIT and Harvard, Cambridge, MA, United States; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States.
Methods Enzymol. 2018;610:191-218. doi: 10.1016/bs.mie.2018.09.019. Epub 2018 Oct 19.
Many promising therapeutic protein targets were previously considered "undruggable" due to a deficit in structural information to guide drug design and/or a lack of an obvious binding pocket. Fortunately, array-based methods for evaluating protein binding against large chemical libraries, such as small-molecule microarray screening, have provided one of several emerging inroads to ligand discovery for these elusive targets. Despite the advance in the area of ligand discovery for poorly structured and intrinsically disordered proteins provided by array-based technologies involving cell lysates, the extension of this technology for screening proteins with short half-lives in physiologically relevant conformations has been technically challenging. In this chapter we present a protocol for leveraging in vitro translation strategies to enable array-based screening of short-lived proteins against large small-molecule libraries for ligand discovery.
许多有前景的治疗性蛋白质靶点以前被认为是“不可成药的”，这是由于缺乏指导药物设计的结构信息和/或缺乏明显的结合口袋。幸运的是，基于阵列的评估蛋白质与大型化学文库结合的方法，如小分子微阵列筛选，为这些难以捉摸的靶点的配体发现提供了几种新兴途径之一。尽管基于细胞裂解物的阵列技术在结构不良和内在无序蛋白质的配体发现领域取得了进展，但将该技术扩展用于筛选具有生理相关构象的短半衰期蛋白质在技术上具有挑战性。在本章中，我们介绍了一种利用体外翻译策略的方案，以实现针对大型小分子文库对短寿命蛋白质进行基于阵列的筛选，用于配体发现。
