Figure The coupling of Stachel-activated GPR56 and disease associated mutations of the receptor with Gα₁₂.

GPR56 or BFPP mutants of the receptor were co-expressed with Gα₁₃ and GRK-based Gβδ biosensors to assess the effect of each mutation on the Gα₁₃ protein coupling, upon receptor activation with the Stachel peptide. For this, HEK293 cells were treated with the 1mM P7 Stachel peptide and Gα₁₃ coupling and heterotrimeric G protein activation was measured as nanoBRET. Under the biosensor conditions reported in this work, all BFPP mutants showed disrupted Gα₁₃coupling while the most severe was observed for GPR56R38W.