Chigaev Alexandre, Sklar Larry A
Department of Pathology and Cancer Center, University of New Mexico Health Sciences Center, Albuquerque, NM, USA.
Methods Mol Biol. 2012;757:3-14. doi: 10.1007/978-1-61779-166-6_1.
Interaction of the integrin receptors with ligands determines the molecular basis of integrin-dependent cell adhesion. Integrin ligands are typically large proteins with relatively low binding affinities. This makes direct ligand-binding kinetic measurements somewhat difficult. Here we examine several real-time methods, aimed to overcome these experimental limitations and to distinguish the regulation of integrin conformation and affinity. This chapter includes: the use of a small ligand-mimetic probe for studies of inside-out regulation of integrin affinity and unbending, real-time cell aggregation and disaggregation kinetics to probe integrin conformational states and the number of integrin-ligand bonds, as well as the real-time monitoring of ligand-induced epitopes under signaling through G-protein-coupled receptors, and others. Experimental data obtained using these novel methods are summarized in terms of the current model of integrin activation.
整合素受体与配体的相互作用决定了整合素依赖性细胞黏附的分子基础。整合素配体通常是具有相对低结合亲和力的大蛋白。这使得直接进行配体结合动力学测量有些困难。在此，我们研究了几种实时方法，旨在克服这些实验限制并区分整合素构象和亲和力的调节。本章包括：使用一种小的配体模拟探针来研究整合素亲和力的外向内调节和伸直，实时细胞聚集和解聚动力学以探测整合素构象状态和整合素 - 配体键的数量，以及在通过G蛋白偶联受体的信号传导下对配体诱导表位的实时监测等。使用这些新方法获得的实验数据根据当前的整合素激活模型进行了总结。
