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. Author manuscript; available in PMC: 2022 Oct 1.

Published in final edited form as: Circ Arrhythm Electrophysiol. 2021 Sep 30;14(10):e010082. doi: 10.1161/CIRCEP.121.010082

Figure 3. — Structural and functional properties of wild-type and mutant W234R G_sα recombinant proteins. A, Wild-type G_sα is protected from trypsin proteolysis after incubation with GTPγS (43kDa proteolytic fragments accumulate) but not with GTP. B, Left panel. Recombinant wild-type and mutant G_sα proteins were incubated with ³⁵[S]GTPγS and analyzed as described in Methods. No effect of mutation on GTP binding was observed and similar K_d values were calculated for both proteins (P=0.46). B, Right panel. Wild-type and mutant G_sα proteins were incubated with [γ−³²P]GTP. Aliquots (50 μl) were analyzed for [³²P]Pi at the indicted time. k_cat was calculated by observation of a single round of GTP hydrolysis performed in triplicate experiments.

Figure 3. — Structural and functional properties of wild-type and mutant W234R G_sα recombinant proteins. A, Wild-type G_sα is protected from trypsin proteolysis after incubation with GTPγS (43kDa proteolytic fragments accumulate) but not with GTP. B, Left panel. Recombinant wild-type and mutant G_sα proteins were incubated with ³⁵[S]GTPγS and analyzed as described in Methods. No effect of mutation on GTP binding was observed and similar K_d values were calculated for both proteins (P=0.46). B, Right panel. Wild-type and mutant G_sα proteins were incubated with [γ−³²P]GTP. Aliquots (50 μl) were analyzed for [³²P]Pi at the indicted time. k_cat was calculated by observation of a single round of GTP hydrolysis performed in triplicate experiments.