Serine Hydrolase Activity‐Based Probes for Use in Chemical Proteomics

Abstract

Serine hydrolases (SHs) comprise a large superfamily of enzymes that play critical roles in many biological processes. Despite their importance, many SHs remain uncharacterized and the vast majority of SHs lack selective inhibitors. In response, activity-based protein profiling (ABPP) and activity-based probes (ABPs) have been leveraged to construct a more comprehensive picture of the SH proteome. Since the utility of ABPP is largely dictated by the reactivity profile of the ABPs deployed, novel scaffolds and chemotypes are needed to expand the breadth and selectivity of SH-targeting ABPs. In this review, we highlight recent innovations in SH probe development, covering both established and emerging electrophilic warheads. We then discuss how strategic implementation of SH-targeting ABPs has yielded selective, potent inhibitors and imaging agents with broad use. Finally, we present methods for ABP diversification and explore cutting-edge applications in therapeutics development and discovery biology.

Emerging and Re-emerging Warheads for Targeted Covalent Inhibitors: An Update

“…Treatment with this molecule led to a nearly complete loss of Ras•GTP and accompanying inhibition of ERK phosphorylation in cancer cell lines. Hydrolysis stability assays in PBS buffer at pH 7.4, however, showed decomposition of half of the compound within 2−4 h. While β-lactones are well known for their (re)activity toward serine hydrolases, 482 their use in the acylation of non-catalytic serine residues had previously not been described, to the best of our knowledge. It is noteworthy that this structural element is present in the approved antiobesity drug orlistat, which has, however, only a very limited systemic availability and that a similar β-lactone-based approach was published by the Shokat group for targeting the aspartate in KRAS G12D while this article was in the proof stage.…”

Lysophosphatidylserine: A Signaling Lipid with Implications in Human Diseases

“…Yet, virtually nothing is known about the biosynthetic or catabolic pathways that regulate lyso-PS metabolism in these tissues. In humans, a majority of the phospholipases and lysophospholipases belong to the mSH family of enzymes, and a significant portion of this enzyme family still lacks the assignment of a physiologically relevant biochemical activity and an in vivo biological function. , Leveraging the repertoire of pharmacological and genetic tools available to interrogate the enzymes of the mSH family, ,− it is possible to determine if certain tissue-specific lipases might, in fact, be metabolic regulators of lyso-PS. For example, the major lyso-PS lipase ABHD12 is highly expressed in the brain and is known to regulate levels of lyso-PS only in the nervous system.…”