Abstract：
Many plasma membrane receptors and ligands form nanoscale clusters on the plasma membrane surface. However, methods for directly and precisely manipulating nanoscale protein localization are limited, making understanding the effects of this clustering difficult. DNA origami allows precise control over nanoscale protein localization with high fidelity and adaptability. Here, we describe how we have used this technique to study how nanoscale protein clustering affects phagocytosis. We provide protocols for conjugating DNA origami structures to supported lipid bilayer-coated beads to assay phagocytosis and planar glass coverslips for TIRF microscopy. The core aspects of this protocol can be translated to study other immune signaling pathways and should enable the implementation of previously inaccessible investigations.
摘要：
许多质膜受体和配体在质膜表面上形成纳米级簇。然而,直接和精确地操纵纳米级蛋白质定位的方法是有限的，这使得理解这种聚类的影响变得困难。DNA折纸允许精确控制纳米级蛋白质定位，具有高保真度和适应性。这里,我们描述了我们如何使用这种技术来研究纳米级蛋白质聚类如何影响吞噬作用。我们提供了将DNA折纸结构缀合到支持的脂质双层涂层珠子上的方案，以测定吞噬作用和用于TIRF显微镜的平面玻璃盖玻片。该协议的核心方面可以转化为研究其他免疫信号通路，并且应该能够实施以前无法进入的调查。
