de Vink Pim J, Andrei Sebastian A, Higuchi Yusuke, Ottmann Christian, Milroy Lech-Gustav, Brunsveld Luc
Laboratory of Chemical Biology , Department of Biomedical Engineering and Institute for Complex Molecular Systems , Eindhoven University of Technology , P. O. Box 513 , 5600MB , Eindhoven , The Netherlands . Email:
The Institute of Scientific and Industrial Research , Osaka University , Ibaraki , Japan.
Chem Sci. 2019 Jan 25;10(10):2869-2874. doi: 10.1039/c8sc05242e. eCollection 2019 Mar 14.
A cooperativity framework to describe and interpret small-molecule stabilization of protein-protein interactions (PPI) is presented. The stabilization of PPIs is a versatile and emerging therapeutic strategy to target specific combinations of protein partners within the protein interactome. Currently, the potency of PPI stabilizers is typically expressed by their apparent affinity or EC. Here, we propose that the effect of a PPI stabilizer be best described involving the cooperativity factor, , between the stabilizer and binding partners in addition to the intrinsic affinity, , of the stabilizer for one of the apo-proteins. By way of illustration, we combine fluorescence polarization measurements with thermodynamic modeling to determine the and for the PPI stabilization of 14-3-3 and TASK3 by fusicoccin-A (FC-A) and validate our approach by studying other PPI-partners of 14-3-3 proteins. Finally, we characterize a library of different stabilizer compounds, and perform structure-activity relationship studies in which molecular changes could be attributed to either changes in cooperativity or intrinsic affinity. Such insights should aid in the development of more effective protein-protein stabilizer drugs.
本文提出了一个用于描述和解释蛋白质-蛋白质相互作用（PPI）小分子稳定作用的协同框架。PPI的稳定作用是一种通用且新兴的治疗策略，旨在靶向蛋白质相互作用组内特定的蛋白质伙伴组合。目前，PPI稳定剂的效力通常用其表观亲和力或EC来表示。在此，我们提出，除了稳定剂对一种无配体蛋白质的固有亲和力Kd之外，PPI稳定剂的作用最好通过稳定剂与结合伙伴之间的协同因子α来描述。作为示例，我们将荧光偏振测量与热力学建模相结合，以确定藤霉素A（FC-A）对14-3-3和TASK3的PPI稳定作用的α和Kd，并通过研究14-3-3蛋白的其他PPI伙伴来验证我们的方法。最后，我们对不同稳定剂化合物库进行了表征，并进行了构效关系研究，其中分子变化可归因于协同性或固有亲和力的变化。这些见解应有助于开发更有效的蛋白质-蛋白质稳定剂药物。
