在细胞中研究Arf GAPs的方法：使用分离的粘着斑进行体外测定。
Approaches to studying Arf GAPs in cells: in vitro assay with isolated focal adhesions.
作者信息
Chen Pei-Wen, Jian Xiaoying, Luo Ruibai, Randazzo Paul A
机构信息
Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland.
出版信息
Curr Protoc Cell Biol. 2012 Jun;Chapter 17:17.13.1-17.13.20. doi: 10.1002/0471143030.cb1713s55.
Abstract
The Arf GAPs are a family of proteins with a common catalytic function of hydrolyzing GTP bound to ADP-ribosylation factors (Arf) with proposed cellular functions that are diverse (Inoue and Randazzo, 2007; Kahn et al., 2008). Understanding the biochemistry of the Arf GAPs is valuable for designing and interpreting experiments using standard cell biology techniques described elsewhere. The following briefly reviews some common approaches for in vivo studies of Arf GAPs and discusses the use of isolated cellular organelles to complement in vivo experiments. Detailed protocols for examining the activity of Arf GAPs in whole cell lysates and in association with isolated focal adhesions are provided.
摘要
Arf GAP蛋白家族具有共同的催化功能，即水解与ADP核糖基化因子（Arf）结合的GTP，其在细胞中的功能多样（井上和兰达佐，2007年；卡恩等人，2008年）。了解Arf GAP蛋白的生物化学特性对于设计和解释使用其他地方所述标准细胞生物学技术的实验很有价值。以下简要回顾了一些用于Arf GAP蛋白体内研究的常见方法，并讨论了使用分离的细胞器来补充体内实验。还提供了检测全细胞裂解物中以及与分离的粘着斑相关的Arf GAP蛋白活性的详细方案。
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