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

The effect of BFPP mutations on Gα13coupling

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.