Fig. 1: Schematic overview of the investigated micro-switch mutants in FZD5 and validation of mutants. | Nature Communications

Fig. 1: Schematic overview of the investigated micro-switch mutants in FZD5 and validation of mutants.

From: Pathway selectivity in Frizzleds is achieved by conserved micro-switches defining pathway-determining, active conformations

Fig. 1

a Design of state-stabilizing mutations. Inactive and active state structures were compared using two GPCRdb tools32 (see Methods) to identify state-specific residue-residue contacts formed by state determinant residues (Supplementary Data 1). b Snake plot of human FZD5 with mutated amino acid residues highlighted in gray. The N and C termini of FZD5 were omitted for clarity. c Validation of cell surface expression in HEK293A cells transiently transfected with the different FZD5 micro-switch mutants, wild-type FZD5, or pcDNA3.1 (Mock) quantified by whole-cell ELISA using an antibody against the N-terminal HA tag. Data show mean ± SEM of six independent experiments performed in triplicate and mean values were normalized to wild-type FZD5 surface expression. Results were analyzed with one-way ANOVA (matched, with Geisser-Greenhouse correction) and uncorrected Fisher’s LSD post-hoc test. Note that only mutants labeled in red are not significantly different from pcDNA3.1 control. Corresponding significance levels are shown in Supplementary Table S2. C-term C terminus, ECL extracellular loop, ICL intracellular loop, N-term N terminus, TM transmembrane domain.

Back to article page