Replacement of the α5 helix of Gα16 with Gαs-specific sequences enhances promiscuity of Gα16 toward Gs-coupled receptors

Characterization of CHO Cells Stably Expressing a Gα16/zChimera for High Throughput Screening of GPCRs

“…11,14 However, as G␣ 16 cannot couple to all GPCRs, 12,17 we have developed and characterized chimeric proteins based on G␣ 16 that incorporate elements of G␣ z , G␣ i , G␣ o , and G␣ s proteins. [18][19][20] Of particular interest are the 16z25 and 16z44 chimeras, which incorporate the C-terminal 25 and 44 amino acids of G␣ z into G␣ 16 (Fig. 1).…”

“…1), 18 as well as a range of chimeras incorporating elements of G␣ s and G␣ i/o . 19,20 These chimeras have proven to recognize Ͼ20 G i/o -and G s -coupled receptors in both FLIPR and reporter gene-based assays using cells transiently coexpressing the receptors and chimeric G proteins. At least 12 of those GPCRs do not demonstrate a substantial functional interaction with G␣ 16 .…”

“…At least 12 of those GPCRs do not demonstrate a substantial functional interaction with G␣ 16 . [17][18][19][20][21] To facilitate the use of these chimeric G proteins in high throughput FLIPR assays, we now describe the generation and characterization of cell lines stably expressing 16z25 and one of nine different GPCRs. 16 (81482) was purchased from Gramsch Laboratories (Schwabhausen, Germany).…”

A Hematopoietic Perspective on the Promiscuity and Specificity of Gα₁₆ Signaling

“…Substitutions of the C-terminal residues in the ␣ 5 helix of G ␣ 16 with those of either G ␣ i2 , G ␣ o1 or G ␣ z can greatly enhance the coupling with G i -linked receptors that normally do not interact with G ␣ 16 [23,26] . The same strategy worked equally well for G s -coupled receptors [27] . Chimeric G ␣ 16 subunits with broadened receptor-coupling capability have been successfully employed in high throughput screening platforms [23,[28][29][30] .…”

Prostacyclin receptor-induced STAT3 phosphorylation in human erythroleukemia cells is mediated via Gαs and Gα16 hybrid signaling

“…The ability of GPCRs to signal via multiple G proteins is fast becoming a rule, and the plethora of G protein signals are often integrated for the regulation of effectors (Lowes et al, 2002 proteins (Mapara et al, 1995;Wilkie et al, 1991 The ability of hIP to signal via the promiscuous Gα 16 (Hazari et al, 2004;Mody et al, 2000;Offermanns and Simon, 1995) is not entirely surprising. Likewise, its poor interaction with Gα 14 is consistent with the less promiscuous nature of Gα 14 (Ho et al, 2001).…”