Extended Data Fig. 3: Construct design and purification of active-state GPR56 and LPHN3 in complex with miniG13 protein.
From: The tethered peptide activation mechanism of adhesion GPCRs
a, Design for the tethered agonist complex constructs used in the study. Receptors are presented on top, where sequences corresponding to the TA and 7TM regions of GPR56 and LPHN3 were inserted after a hemagglutinin signal peptide (HA) and a methionine residue. Expression vector for the miniG13 heterotrimer presented at the bottom. b, The MiniGα13/i15 sequence. Residues at the N-terminus corresponding to Gαi2 sequence are in grey. The linker replacing the alpha helical domain is in yellow. Residues corresponding to the stabilizing mutations G57DS1H1.03, E58NS1H1.04, S248DS4.07, E251DS4H3.03, I271DH3.08, I355AH5.04, V358IH5.07 are underlined and presented in bold. c, Size-exclusion chromatography (SEC) profiles of purified miniG13-coupled GPR56 (left) and -LPHN3 (right) with insets showing Coomassie-stained SDS-PAGE of the SEC complex peaks.
