Department of Chemistry, Wayne State University, 5101 Cass Avenue, Detroit, MI 48202, USA.
Biotechniques. 2018 Sep;65(3):143-148. doi: 10.2144/btn-2018-0076.
Kinases are essential cell signaling enzymes that phosphorylate protein substrates using ATP as the universal cosubstrate. A wide variety of ATP analogs have been used in kinase research, although the studies are limited by the cell impermeability of ATP. Here we describe the use of the cationic polymer deacetylated chitosan to permeabilize ATP analogs for live cell applications, including kinase-catalyzed biotinylation.
激酶是细胞信号转导中必不可少的酶，它们使用 ATP 作为通用辅助底物来磷酸化蛋白质底物。在激酶研究中已经使用了多种 ATP 类似物，尽管这些研究受到 ATP 细胞通透性的限制。在这里，我们描述了使用阳离子聚合物脱乙酰壳聚糖来透化 ATP 类似物，以应用于活细胞，包括激酶催化的生物素化。
