Proteome-Wide Reactivity Profiling Identifies Diverse
Carbamate Chemotypes Tuned for Serine Hydrolase Inhibition
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Abstract
Serine
hydrolases are one of the largest and most diverse enzyme
classes in Nature. Inhibitors of serine hydrolases are used to treat
many diseases, including obesity, diabetes, cognitive dementia, and
bacterial and viral infections. Nonetheless, the majority of the 200+
serine hydrolases in mammals still lack selective inhibitors for their
functional characterization. We and others have shown that activated
carbamates, through covalent reaction with the conserved serine nucleophile
of serine hydrolases, can serve as useful inhibitors for members of
this enzyme family. The extent to which carbamates, however, cross-react
with other protein classes remains mostly unexplored. Here, we address
this problem by investigating the proteome-wide reactivity of a diverse
set of activated carbamates <i>in vitro</i> and <i>in vivo</i>, using a combination of competitive and click chemistry
(CC)-activity-based protein profiling (ABPP). We identify multiple
classes of carbamates, including <i>O</i>-aryl, <i>O</i>-hexafluoroisopropyl (HFIP), and <i>O</i>-<i>N</i>-hydroxysuccinimidyl (NHS) carbamates that react selectively
with serine hydrolases across entire mouse tissue proteomes <i>in vivo</i>. We exploit the proteome-wide specificity of HFIP
carbamates to create <i>in situ</i> imaging probes for the
endocannabinoid hydrolases monoacylglycerol lipase (MAGL) and α-β
hydrolase-6 (ABHD6). These findings, taken together, designate the
carbamate as a privileged reactive group for serine hydrolases that
can accommodate diverse structural modifications to produce inhibitors
that display exceptional potency and selectivity across the mammalian
proteome