Fig. 3: ATGL switch mutations selectively interrupt individual binding partners.
From: Mutational scanning pinpoints distinct binding sites of key ATGL regulators in lipolysis
A Luciferase-based co-immunoprecipitation of 52 ATGL mutants with five protein interaction partners. Box plot of showing the mean of triplicate measurements of two experiments each probing murine Plin5 (blue), murine Plin1 (cyan), G0S2 (yellow), CIDEC (red), and CGI-58 (green) for interaction with the human ATGL variants (gray data points), ATGL wild-type (green data points) and a negative control (red data points). Log2 fold changes in binding relative to ATGL wild-type was determined within every experiment (Supplementary Data 4). Values were normalized across the experiments to the 3rd quartile indicating increased or decreased interaction signal. The boxes extend from 1st to 3rd quartile with the central band representing the median. The whiskers extend to the furthest point up to 1.5 times the interquartile range away from the nearest quartile. B Systematic overview of the binding data of the 52 ATGL variants. Variants are shown with their location within the patatin domain or in the C-terminal half of the protein. ATGL variants reducing the binding by more than twofold are marked as colored dots according to the affected interaction partner. The switch mutants are highlighted separately. Color code: CGI-58 green, CIDEC red, G0S2 yellow, mPlin1 cyan and mPlin5 blue. C Binding data for eleven ATGL switch mutations. For each interaction partner (color coded) normalized log2FC of two experiments performed in triplicates are shown. Single amino acid ATGL variants are color-coded according to the most specific binding alteration with a single partner, color code: CGI-58 green, CIDEC red, G0S2 yellow, mPlin1 cyan, and mPlin5 blue. Log2FC values within −0.5–0.5 (gray area) indicate wild-type-like binding behavior.
