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. Author manuscript; available in PMC: 2020 Dec 8.
Published in final edited form as: Nat Chem Biol. 2020 Jun 8;16(9):997–1005. doi: 10.1038/s41589-020-0555-4

Figure 4. Optimization of LYPLAL1 activators.

Figure 4.

(a) Representative analogues of 4 that display greater potency and efficacy on LYPLAL1 overexpressed in HEK293 cells, as assessed by gel-based ABPP. Western blots of LYPLAL1 and HSP90 (loading control) are shown in the bottom panels. (b) Endogenous LYPLAL1 is selectively activated in total HepG2 proteomes treated with 10 μM 12 (or PAL-12) for 1 h and then labeled with FP-alkyne for 20 min prior to attachment of an azide-biotin tag and ABPP-MudPIT analysis (n=2, where n represents independent samples). (c) Thermal stability profiles of purified mLYPLAL1 (at 49°C) show that interaction with increasing concentrations of 4 and 12 increase the protein’s flexibility (n=3 per group, where n represents independent experiments). Error bars represent mean ± s.d. Representative results from two (b) or three (c) independent experiments; similar results were obtained in all experiments. Uncropped gels/blots for a are shown in Supplementary Fig. 6.