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.

中文翻译：

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一种功能中性的单链抗体，可测量β-1整联蛋白的摄取和再循环。

整联蛋白介导的细胞粘附和信号传导对于许多生理过程至关重要。整联蛋白及其相关的粘附复合物通过内吞和再循环途径的动态转换已经成为控制癌症中细胞迁移和侵袭的重要机制。因此，对整联蛋白运输的调控以及疾病特异性分子机制如何改变这种调控引起了极大的兴趣。但是，当前可用于研究整联蛋白运输的工具可能会导致伪影和/或无法提供足够的动力学信息。在这里，我们报告功能性中性和单价单链抗体的生成，以定量和定性地测量细胞中β1整联蛋白的运输。我们的新型探针可用于多种测定中，可对内源性整联蛋白进行快速回收利用，进行生物化学表征。我们还展示了其在活细胞成像中的潜在效用，为指导未来整联蛋白探针设计提供了原理证明。