使用荧光MANT-GDP的鸟嘌呤核苷酸交换测定法。
Guanine Nucleotide Exchange Assay Using Fluorescent MANT-GDP.
作者信息
Kanie Tomoharu, Jackson Peter K
机构信息
Baxter Laboratory, Department of Microbiology & Immunology and Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA.
出版信息
Bio Protoc. 2018 Apr 5;8(7). doi: 10.21769/BioProtoc.2795.
Abstract
GTPases are molecular switches that cycle between the inactive GDP-bound state and the active GTP-bound state. GTPases exchange nucleotides either by its intrinsic nucleotide exchange or by interaction with guanine nucleotide exchange factors (GEFs). Monitoring the nucleotide exchange , together with reconstitution of direct interactions with regulatory proteins, provides key insights into how a GTPase is activated. In this protocol, we describe core methods to monitor nucleotide exchange using fluorescent N-Methylanthraniloyl (MANT)-guanine nucleotide.
摘要
GTP酶是在无活性的结合GDP状态和有活性的结合GTP状态之间循环的分子开关。GTP酶通过其内在的核苷酸交换或与鸟嘌呤核苷酸交换因子（GEFs）相互作用来交换核苷酸。监测核苷酸交换，以及与调节蛋白直接相互作用的重组，为深入了解GTP酶如何被激活提供了关键见解。在本实验方案中，我们描述了使用荧光N-甲基邻氨基苯甲酰（MANT）-鸟嘌呤核苷酸监测核苷酸交换的核心方法。
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