Abstract：
Ras and Rap small GTPases of the Ras superfamily act as molecular switches to control diverse cellular processes as part of different signaling pathways. Dictyostelium expresses several Ras and Rap proteins, and their study has and continues to greatly contribute to our understanding of their role in eukaryote biology. To study the activity of Ras and Rap proteins in Dictyostelium, several assays based on their interaction with the Ras binding domain of known eukaryotic Ras/Rap effectors have been developed and proved extremely useful to study their regulation and cellular roles. Here, we describe methods to assess Ras/Rap activity biochemically using a pull-down assay and through live-cell imaging using fluorescent reporters.
摘要：
Ras超家族的Ras和Rap小GTP酶作为分子开关来控制作为不同信号传导途径的一部分的不同细胞过程。Dictyostelium表达几种Ras和Rap蛋白，他们的研究已经并将继续大大有助于我们理解他们在真核生物中的作用。为了研究网菌属Ras和Rap蛋白的活性，已经开发了几种基于它们与已知真核Ras/Rap效应子的Ras结合域相互作用的测定法，并证明对研究它们的调节和细胞作用非常有用。这里,我们描述了使用下拉测定法和使用荧光报道分子通过活细胞成像生化评估Ras/Rap活性的方法。
