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. 2018 Jul 26;1(1):12–20. doi: 10.1021/acsptsci.8b00017

Figure 2.

Figure 2

Comparison of purification of WT-Gs and DN-Gs containing heterotrimers with CTR and GLP-1R. (a) Coomassie stained gel showing relative abundance of various components of CTR (with salmon calcitonin) and GLP-1R (with exendin-P5)–G protein complexes following FLAG purification in the presence of either WT or DN-Gs but not Nb35. (b) 2D class averages from negative stained EM micrographs of the CTR:DN Gs and GLP-1R:DN Gs complexes in the absence of Nb35. (c) Western blot against His-tag (Gβ, Nb35, and CTR:GLP-1R) and Gαs from purified active complex with DN-Gs in the presence or absence of Nb35. (d,e), Western blot and coomassie stained gel of WT- (d) and DN- (e) containing GLP-1R:Gs complexes in the presence of Nb35 and corresponding SEC traces (f), illustrating that both complexes can be purified but that the proportion of complex in the void/aggregate is slightly higher with the WT-Gαs (c).