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a <t>GTP</t> <t>hydrolysis</t> of the S4 mediated Gα s βγ as a function of inverse, partial, and full agonists, in reference to Gα s βγ alone (negative control) and the S5 mediated Gα s βγ (positive control). Data with error bars are presented as mean ± SEM of four independent experiments. Statistical analyses were performed using the ordinary one-way ANOVA compared to the R291A apo, *** p < 0.001, and **** p < 0.0001. b Time course of GTP hydrolysis of the S4 mediated Gα s βγ, in reference to Gα s βγ alone (negative control) and the S5 mediated Gα s βγ (positive control); the initial rate of each catalysis was calculated. The initial rate of each catalysis was calculated. Data are presented as mean values ± SD from three independent experiments ( n = 3). Source data are provided in the Source Data file. c Km values for S4 mediated Gα s βγ, in reference to Gα s βγ alone (negative control) and the S5 mediated Gα s βγ (positive control). Data are presented as mean values ± SD from three independent experiments ( n = 3). Source data are provided in the Source Data file. d BODIPY-FL-GDP binding assay. e BODIPY-FL-GTP binding assay. f BODIPY-FL-GTP-γ-S binding assay ( g ) GTP-GDP exchange rate comparison. h The structures of BODIPY-FL-GDP, BODIPY - FL-GTP, and BODIPY-FL-GTP-γ-S.
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a GTP hydrolysis of the S4 mediated Gα s βγ as a function of inverse, partial, and full agonists, in reference to Gα s βγ alone (negative control) and the S5 mediated Gα s βγ (positive control). Data with error bars are presented as mean ± SEM of four independent experiments. Statistical analyses were performed using the ordinary one-way ANOVA compared to the R291A apo, *** p < 0.001, and **** p < 0.0001. b Time course of GTP hydrolysis of the S4 mediated Gα s βγ, in reference to Gα s βγ alone (negative control) and the S5 mediated Gα s βγ (positive control); the initial rate of each catalysis was calculated. The initial rate of each catalysis was calculated. Data are presented as mean values ± SD from three independent experiments ( n = 3). Source data are provided in the Source Data file. c Km values for S4 mediated Gα s βγ, in reference to Gα s βγ alone (negative control) and the S5 mediated Gα s βγ (positive control). Data are presented as mean values ± SD from three independent experiments ( n = 3). Source data are provided in the Source Data file. d BODIPY-FL-GDP binding assay. e BODIPY-FL-GTP binding assay. f BODIPY-FL-GTP-γ-S binding assay ( g ) GTP-GDP exchange rate comparison. h The structures of BODIPY-FL-GDP, BODIPY - FL-GTP, and BODIPY-FL-GTP-γ-S.

Journal: Nature Communications

Article Title: Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex

doi: 10.1038/s41467-025-56434-4

Figure Lengend Snippet: a GTP hydrolysis of the S4 mediated Gα s βγ as a function of inverse, partial, and full agonists, in reference to Gα s βγ alone (negative control) and the S5 mediated Gα s βγ (positive control). Data with error bars are presented as mean ± SEM of four independent experiments. Statistical analyses were performed using the ordinary one-way ANOVA compared to the R291A apo, *** p < 0.001, and **** p < 0.0001. b Time course of GTP hydrolysis of the S4 mediated Gα s βγ, in reference to Gα s βγ alone (negative control) and the S5 mediated Gα s βγ (positive control); the initial rate of each catalysis was calculated. The initial rate of each catalysis was calculated. Data are presented as mean values ± SD from three independent experiments ( n = 3). Source data are provided in the Source Data file. c Km values for S4 mediated Gα s βγ, in reference to Gα s βγ alone (negative control) and the S5 mediated Gα s βγ (positive control). Data are presented as mean values ± SD from three independent experiments ( n = 3). Source data are provided in the Source Data file. d BODIPY-FL-GDP binding assay. e BODIPY-FL-GTP binding assay. f BODIPY-FL-GTP-γ-S binding assay ( g ) GTP-GDP exchange rate comparison. h The structures of BODIPY-FL-GDP, BODIPY - FL-GTP, and BODIPY-FL-GTP-γ-S.

Article Snippet: All statistical tests such as GTP hydrolysis assessments were conducted using GraphPad Prism 9.0.

Techniques: Negative Control, Positive Control, Binding Assay, GTP Binding Assay, Comparison