Fig. 1: Degradation of SHP2 by E2D1-based bioPROTACs.
From: Targeted protein degradation using chimeric human E2 ubiquitin-conjugating enzymes
a Proposed intracellular biological mechanism for E2 bioPROTAC activity. Ubiquitin is first captured by the E2 enzymatic domain (step 1). The target protein is recruited via the attached binding domain [BD] and subsequently ubiquitinated (step 2). Re-loading of the E2 bioPROTAC (step 3) enables further target ubiquitination, ultimately leading to its degradation in the proteasome (step 4). Created with BioRender.com. b Immunofluorescence (IF) images of U2OS cells fixed 24 h after transfection with bioPROTAC or isotype control mRNAs. Scale bar (lower right corner image) = 100 µm. c, d Dot plots showing mean cellular SHP2 levels and expressed HA-tagged protein levels from IF images in (b). Data from three biological replicates. e Representative western blot of U2OS cells transfected with SHP2-targeted bioPROTAC or isotype control mRNAs and harvested after 24 h. f Mean SHP2 densitometry data from (e) (n = 3 biological replicates). Statistical significance for data in (c, d, f) was calculated using a one-way ANOVA.
