细胞裂解物中活化的GAP和GEF分析。

Analysis of activated GAPs and GEFs in cell lysates.

作者信息

García-Mata Rafael, Wennerberg Krister, Arthur William T, Noren Nicole K, Ellerbroek Shawn M, Burridge Keith

机构信息

Department of Cell and Developmental Biology, Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, USA.

出版信息

Methods Enzymol. 2006;406:425-37. doi: 10.1016/S0076-6879(06)06031-9.

DOI:10.1016/S0076-6879(06)06031-9

PMID:16472675

Abstract

An assay was developed that allows the precipitation of the active pools of Rho-GEFs, Rho-GAPs, or effectors from cell or tissue lysates. This assay can be used to identify GEFs, GAPs, and effectors involved in specific cellular pathways to determine their GTPase specificity and to monitor the temporal activation of GEFs and GAPs in response to upstream signals.

摘要

开发了一种检测方法，可从细胞或组织裂解物中沉淀出活性池中的Rho鸟嘌呤核苷酸交换因子（Rho-GEFs）、Rho鸟嘌呤核苷酸酶激活蛋白（Rho-GAPs）或效应器。该检测方法可用于识别参与特定细胞通路的鸟嘌呤核苷酸交换因子、鸟嘌呤核苷酸酶激活蛋白和效应器，以确定它们的GTP酶特异性，并监测鸟嘌呤核苷酸交换因子和鸟嘌呤核苷酸酶激活蛋白对上游信号响应的瞬时激活。