Tang Tuo-Xian, Xiong Wen, Finkielstein Carla V, Capelluto Daniel G S
Protein Signaling Domains Laboratory, Department of Biological Sciences, Biocomplexity Institute, Virginia Tech, Blacksburg, VA, 24061, USA.
Center for Soft Matter and Biological Physics, Virginia Tech, Blacksburg, VA, 24061, USA.
Methods Mol Biol. 2017;1647:197-206. doi: 10.1007/978-1-4939-7201-2_13.
The protein-lipid overlay assay is an inexpensive, easy-to-implement, and high-throughput methodology that employs nitrocellulose membranes to immobilize lipids in order to rapid screen and identify protein-lipid interactions. In this chapter, we show how this methodology can identify potential modulators of protein-lipid interactions by screening water-soluble lipid competitors or even the introduction of pH changes during the binding assay to identify pH-dependent lipid binding events.
蛋白质-脂质覆盖分析是一种廉价、易于实施且高通量的方法，该方法利用硝酸纤维素膜固定脂质，以便快速筛选和鉴定蛋白质-脂质相互作用。在本章中，我们展示了这种方法如何通过筛选水溶性脂质竞争剂，甚至在结合分析过程中引入pH变化以识别pH依赖性脂质结合事件，来鉴定蛋白质-脂质相互作用的潜在调节剂。
