Department of Pathology, University of New Mexico School of Medicine, Albuquerque, NM 87131, USA; Cancer Center, University of New Mexico School of Medicine, Albuquerque, NM 87131, USA; Center for Infectious Diseases and Immunity, University of New Mexico School of Medicine, Albuquerque, NM 87131, USA.
Anal Biochem. 2013 Nov 15;442(2):149-57. doi: 10.1016/j.ab.2013.07.039. Epub 2013 Aug 6.
We describe a rapid assay for measuring the cellular activity of small guanine triphosphatases (GTPases) in response to a specific stimulus. Effector-functionalized beads are used to quantify in parallel multiple GTP-bound GTPases in the same cell lysate by flow cytometry. In a biologically relevant example, five different Ras family GTPases are shown for the first time to be involved in a concerted signaling cascade downstream of receptor ligation by Sin Nombre hantavirus.
我们描述了一种快速测定法，用于测量细胞中小鸟苷三磷酸酶（GTPases）对特定刺激的反应的细胞活性。效应功能化的珠子通过流式细胞术用于同时定量同一细胞裂解物中的多个结合 GTP 的 GTPase。在一个生物学相关的例子中，首次表明五个不同的 Ras 家族 GTPase 参与了辛诺伯尔病毒受体结合后的协调信号级联反应。
