Wellcome Trust Centre for Cell-Matrix Research, Division of Cell Matrix Biology and Regenerative Medicine, School of Biological Sciences, University of Manchester, Manchester, UK.
Methods Mol Biol. 2021;2217:251-263. doi: 10.1007/978-1-0716-0962-0_14.
The last two decades of research into integrin trafficking has revealed fascinating insight into the function of integrin receptors, particularly in the context of cell invasion and migration in cancer. Deregulation in the trafficking pathways of integrin receptors contributes to a variety of pathological conditions including cancer, and in fact, altered endocytic trafficking of these receptors has been shown to drive transformation and tumor progression. Being able to experimentally measure integrin internalization, recycling and cell surface levels are vital for determining the role integrins play in health and disease. Surface-labeling based endocytic trafficking assays provide a way to experimentally measure changes in the rate of internalization of cell surface proteins, and the recycling of internalized proteins back to the cell surface, with high accuracy. This chapter will focus on quantitative approaches based on cell surface biotinylation and capture ELISA to measure endocytosis, recycling, and cell surface levels of integrin receptors.
过去二十年对整合素运输的研究揭示了整合素受体功能的迷人见解，特别是在癌症中细胞侵袭和迁移的背景下。整合素受体运输途径的失调导致多种病理状况，包括癌症，事实上，这些受体的内吞运输改变已被证明可驱动转化和肿瘤进展。能够通过实验测量整合素内化、回收和细胞表面水平对于确定整合素在健康和疾病中的作用至关重要。基于表面标记的内吞运输测定提供了一种实验测量细胞表面蛋白内化率以及内化蛋白回收到细胞表面的速率变化的方法，具有很高的准确性。本章将重点介绍基于细胞表面生物素化和捕获 ELISA 的定量方法，以测量整合素受体的内吞作用、回收和细胞表面水平。
