Department of Cell Biology, UT Southwestern Medical Center, Dallas, Texas, USA.
Traffic. 2020 Sep;21(9):590-602. doi: 10.1111/tra.12754. Epub 2020 Jul 21.
Integrin-mediated cell adhesion and signaling are critical for many physiological processes. The dynamic turnover of integrins and their associated adhesion complexes through endocytic and recycling pathways has emerged as an important mechanism for controlling cell migration and invasion in cancer. Thus, the regulation of integrin trafficking and how this may be altered by disease-specific molecular mechanisms has generated considerable interest. However, current tools available to study integrin trafficking may cause artifacts and/or do not provide adequate kinetic information. Here, we report the generation of a functionally neutral and monovalent single chain antibody to quantitatively and qualitatively measure β1 integrin trafficking in cells. Our novel probe can be used in a variety of assays and allows for the biochemical characterization of rapid recycling of endogenous integrins. We also demonstrate its potential utility in live cell imaging, providing proof of principle to guide future integrin probe design.
整合素介导的细胞黏附和信号转导对于许多生理过程至关重要。通过内吞和回收途径，整合素及其相关黏附复合物的动态周转已成为控制癌症中细胞迁移和侵袭的重要机制。因此，整合素运输的调节以及疾病特异性分子机制如何改变它引起了相当大的兴趣。然而，目前用于研究整合素运输的工具可能会产生假象和/或不能提供足够的动力学信息。在这里，我们报告了一种功能中性的单价单链抗体的产生，用于定量和定性测量细胞中β1 整合素的运输。我们的新型探针可用于多种检测，并允许对快速内源性整合素的回收进行生化表征。我们还证明了它在活细胞成像中的潜在用途，为指导未来整合素探针设计提供了原理证明。
