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. Author manuscript; available in PMC: 2014 Aug 1.
Published in final edited form as: Nat Chem Biol. 2013 Dec 22;10(2):113–121. doi: 10.1038/nchembio.1429

Figure 1. Isolation of serine hydrolase inhibitors with adipogenic activity and identification of their molecular target.

Figure 1

(a) Hierarchical cluster analysis of serine hydrolase signals detected by ABPP-MudPIT in 3T3-L1 and 10T1/2 preadipocyte and adipocyte proteomes. Data represent the normalized mean of three independent experiments. (b) Gel-based competitive ABPP analysis of adipocyte 10T1/2 cells labeled in situ with carbamates that promote differentiation and lipid accumulation in fat cells. A ~60 kDa serine hydrolase (black arrow) is inhibited by multiple proadipogenic carbamates. WWL38 inhibits HSL (grey arrow); WWL113 appears specific for the 60kDa activity. Image is representative of 2 independent experiments. (c) Structure of WWL113 and its urea derivative (WWL113U). WWL113, but not WWL113U, promotes adipocyte formation and lipid storage in 10T1/2 cells. Green fluorescence corresponds to Nile red staining. Images are representative of 10 independent experiments. Scale bar = 250 μm. (d) Competitive ABPP shows that WWL113 specifically targets a ~60 kDa serine hydrolase activity that is highly induced during differentiation of 10T1/2 adipocytes; WWL113U has no effect. Image is representative of 3 independent experiments. (e) Competitive ABPP-MudPIT analysis of proteomes from 10T1/2 adipocytes incubated with WWL113 reveals that this compound is a Ces3/Ces1f inhibitor. Error bars represent s.d. (n = 3).