Hooks Shelley B, Waldo Gary L, Corbitt James, Bodor Erik T, Krumins Andrejs M, Harden T Kendall
Department of Pharmacology, University of North Carolina, Chapel Hill 27599, USA.
J Biol Chem. 2003 Mar 21;278(12):10087-93. doi: 10.1074/jbc.M211382200. Epub 2003 Jan 16.
Regulator of G-protein signaling (RGS) proteins are GTPase activating proteins (GAPs) of heterotrimeric G-proteins that alter the amplitude and kinetics of receptor-promoted signaling. In this study we defined the G-protein alpha-subunit selectivity of purified Sf9 cell-derived R7 proteins, a subfamily of RGS proteins (RGS6, -7, -9, and -11) containing a Ggamma-like (GGL) domain that mediates dimeric interaction with Gbeta(5). Gbeta(5)/R7 dimers stimulated steady state GTPase activity of Galpha-subunits of the G(i) family, but not of Galpha(q) or Galpha(11), when added to proteoliposomes containing M2 or M1 muscarinic receptor-coupled G-protein heterotrimers. Concentration effect curves of the Gbeta(5)/R7 proteins revealed differences in potencies and efficacies toward Galpha-subunits of the G(i) family. Although all four Gbeta(5)/R7 proteins exhibited similar potencies toward Galpha(o), Gbeta(5)/RGS9 and Gbeta(5)/RGS11 were more potent GAPs of Galpha(i1), Galpha(i2), and Galpha(i3) than were Gbeta(5)/RGS6 and Gbeta(5)/RGS7. The maximal GAP activity exhibited by Gbeta(5)/RGS11 was 2- to 4-fold higher than that of Gbeta(5)/RGS7 and Gbeta(5)/RGS9, with Gbeta(5)/RGS6 exhibiting an intermediate maximal GAP activity. Moreover, the less efficacious Gbeta(5)/RGS7 and Gbeta(5)/RGS9 inhibited Gbeta(5)/RGS11-stimulated GTPase activity of Galpha(o). Therefore, R7 family RGS proteins are G(i) family-selective GAPs with potentially important differences in activities.
G蛋白信号调节(RGS)蛋白是异三聚体G蛋白的GTP酶激活蛋白(GAP),可改变受体介导信号传导的幅度和动力学。在本研究中,我们确定了纯化的Sf9细胞来源的R7蛋白的G蛋白α亚基选择性,R7蛋白是RGS蛋白(RGS6、-7、-9和-11)的一个亚家族,含有一个Gγ样(GGL)结构域,介导与Gβ5的二聚体相互作用。当添加到含有M2或M1毒蕈碱受体偶联的G蛋白异三聚体的蛋白脂质体中时,Gβ5/R7二聚体刺激G(i)家族Gα亚基的稳态GTP酶活性,但不刺激Gαq或Gα11的活性。Gβ5/R7蛋白的浓度效应曲线显示,它们对G(i)家族Gα亚基的效力和效能存在差异。尽管所有四种Gβ5/R7蛋白对Gαo的效力相似,但Gβ5/RGS9和Gβ5/RGS11对Gαi1、Gαi2和Gαi3的GAP活性比Gβ5/RGS6和Gβ5/RGS7更强。Gβ5/RGS11表现出的最大GAP活性比Gβ5/RGS7和Gβ5/RGS9高2至4倍,Gβ5/RGS6表现出中等的最大GAP活性。此外,效力较低的Gβ5/RGS7和Gβ5/RGS9抑制Gβ5/RGS11刺激的Gαo的GTP酶活性。因此,R7家族RGS蛋白是G(i)家族选择性GAP,其活性可能存在重要差异。