Willard Francis S, Siderovski David P
Department of Pharmacology, The University of North Carolina at Chapel Hill, 27599-7365, USA.
Methods Enzymol. 2004;389:320-38. doi: 10.1016/S0076-6879(04)89019-0.
The model organism Arabidopsis thaliana contains a restricted set of heterotrimeric G-protein subunits, with only one canonical Galpha subunit (AtGPA1), one Gbeta subunit (AtAGB1), and two Ggamma subunits (AtAGG1 and AtGG2) identified. We have identified a novel additional component of heterotrimeric G-protein signaling in the A. thaliana genome, regulator of G-protein signaling-1 (AtRGS1). This protein has the predicted topology and structure of a G-protein-coupled receptor in that it contains seven transmembrane domains, but AtRGS1 also contains a unique C-terminal extension, namely a regulator of G-protein signaling domain (RGS box). This article describes methods for the purification and in vitro functional analysis of the RGS box of AtRGS1.
模式生物拟南芥仅包含一组有限的异源三聚体G蛋白亚基，仅鉴定出一个典型的Gα亚基（AtGPA1）、一个Gβ亚基（AtAGB1）和两个Gγ亚基（AtAGG1和AtGG2）。我们在拟南芥基因组中鉴定出了异源三聚体G蛋白信号传导的一个新的额外组分，即G蛋白信号调节剂-1（AtRGS1）。该蛋白具有预测的G蛋白偶联受体的拓扑结构和结构，因为它包含七个跨膜结构域，但AtRGS1还包含一个独特的C末端延伸，即G蛋白信号调节结构域（RGS结构域）。本文描述了AtRGS1的RGS结构域的纯化及体外功能分析方法。
