Fig. 8: BMP10:BMPRII interaction is important for prodomain displacement. | Nature Communications

Fig. 8: BMP10:BMPRII interaction is important for prodomain displacement.

From: Crystal structures of BMPRII extracellular domain in binary and ternary receptor complexes with BMP10

Fig. 8

a A representative native PAGE of prodomain displacement experiment. b Quantification of native PAGE results. The band intensities of Pro:BMP10 and prodomain were obtained by densitometry using ImageJ. The ratio of Pro:BMP10/prodomain for each lane was calculated, then normalised to that of the WT control run on the same gel. Each mutant protein was run on at least three independent native PAGE experiments, with two WT controls on each gel to allow normalisation. Final N numbers for each sample are labelled below the graph. Conditions with BMPRII ECDs at 2-fold excess (2x) are shown in open bars, and those with 5x in hashed bars. The two dotted lines mark the mean values for 2x and 5x WT BMPRII, respectively. Data are presented as mean values + /− SEM. One-way ANOVA analysis for WT ECD group, ****P < 0.0001, **P < 0.01; for BMPRII at 2x excess group, Dunnett’s post test comparing with WT BMPRII ECD, $P < 0.05, $$P < 0.01; for BMPRII at 5x excess group, Dunnett’s post test comparing with WT BMPRII ECD, #P < 0.05, ###P < 0.001, ####P < 0.0001. c Schematic depicting the experiment of the prodomain displacement by ELISA. d Quantitative results of prodomain displacement by ELISA. All readings are normalised to the WT on the same plate. N = 5 for WT ECD, N = 3 for each mutant protein. Each N number represents an independent ELISA experiment. Data are presented as mean values + /− SEM. One-way ANOVA analysis for BMPRII at 125x excess, with Dunnett’s post test comparing with WT BMPRII ECD (mean value shown as a dotted line). #P < 0.05, ##P < 0.01, ###P < 0.001, ####P < 0.0001. In b, d, coloured in red are residues near the hydrophobic triad, in orange are residues from the A-loop, in yellow are F3-loop; in brown, non-interface residues, and in purple, F1-loop which is not visible in the structure. Source data, including the exact p values for b, d, are provided as a Source Data file. ECD = extracellular domain, GF-domain = growth factor domain, Pro:BMP10 = non-covalent complex of BMP10 prodomain with its GF-domain.

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