荧光探针 XY-69 在脂质体中用于测量磷酯酶 C 同工酶的活性。
Fluorogenic XY-69 in Lipid Vesicles for Measuring Activity of Phospholipase C Isozymes.
机构信息
Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
出版信息
Methods Mol Biol. 2021;2251:225-236. doi: 10.1007/978-1-0716-1142-5_17.
Mammalian phospholipase C (PLC) isozymes are major signaling nodes that regulate a wide range of cellular processes. Dysregulation of PLC activity has been associated with a growing list of human diseases such as cancer and Alzheimer's disease. However, methods to directly and continuously monitor PLC activity at membranes with high sensitivity and throughput are still lacking. We have developed XY-69, a fluorogenic PIP analog, which can be efficiently hydrolyzed by PLC isozymes either in solution or at membranes. Here, we describe the optimized assay conditions and protocol to measure the activity of PLC-γ1 (D1165H) with XY-69 in lipid vesicles. The described protocol also applies to other PLC isozymes.
哺乳动物磷酯酶 C（PLC）同工酶是调节广泛细胞过程的主要信号节点。PLC 活性的失调与越来越多的人类疾病有关，如癌症和阿尔茨海默病。然而，直接且持续地以高灵敏度和高通量监测膜上 PLC 活性的方法仍然缺乏。我们开发了 XY-69，一种荧光 PIP 类似物，可在溶液或膜中被 PLC 同工酶有效水解。在此，我们描述了使用 XY-69 在脂质小泡中测量 PLC-γ1（D1165H）活性的优化测定条件和方案。该方案也适用于其他 PLC 同工酶。
Zotero 插件