Gao Xinxin, Hannoush Rami N
Department of Cell Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX, 75390-9039, USA.
Methods Mol Biol. 2016;1481:1-9. doi: 10.1007/978-1-4939-6393-5_1.
Wnt palmitoylation regulates its secretion and signaling activity in cells. Methods to monitor cellular Wnt palmitoylation are instrumental in investigating Wnt activity, secretion, and its interaction with cellular membrane compartments. This protocol describes a method we have recently developed to detect cellular Wnt palmitoylation. The method, combining click chemistry, bio-orthogonal fatty acid probes, and proximity ligation assay (PLA), provides high sensitivity and subcellular resolution for detection of Wnt palmitoylation. It is also compatible with multiple imaging platforms, and is applicable to detecting palmitoylated forms of other fatty acylated proteins.
Wnt蛋白的棕榈酰化调节其在细胞中的分泌和信号活性。监测细胞中Wnt蛋白棕榈酰化的方法对于研究Wnt活性、分泌及其与细胞膜区室的相互作用至关重要。本方案描述了我们最近开发的一种检测细胞中Wnt蛋白棕榈酰化的方法。该方法结合了点击化学、生物正交脂肪酸探针和邻近连接分析(PLA),为检测Wnt蛋白棕榈酰化提供了高灵敏度和亚细胞分辨率。它还与多种成像平台兼容,适用于检测其他脂肪酰化蛋白的棕榈酰化形式。