1. Functional annotation of serine hydrolases in the asexual erythrocytic stage of Plasmodium falciparum.
- Author
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Elahi R, Ray WK, Dapper C, Dalal S, Helm RF, and Klemba M
- Subjects
- Biotin analogs & derivatives, Humans, Hydrolases antagonists & inhibitors, Life Cycle Stages, Lipolysis, Plasmodium falciparum growth & development, Proteomics, Serine metabolism, Erythrocytes parasitology, Hydrolases metabolism, Plasmodium falciparum enzymology, Protozoan Proteins metabolism
- Abstract
Enzymes of the serine hydrolase superfamily are ubiquitous, highly versatile catalysts that mediate a wide variety of metabolic reactions in eukaryotic cells, while also being amenable to selective inhibition. We have employed a fluorophosphonate-based affinity capture probe and mass spectrometry to explore the expression profile and metabolic roles of the 56-member P. falciparum serine hydrolase superfamily in the asexual erythrocytic stage of P. falciparum. This approach provided a detailed census of active serine hydrolases in the asexual parasite, with identification of 21 active serine hydrolases from α/β hydrolase, patatin, and rhomboid protease families. To gain insight into their functional roles and substrates, the pan-lipase inhibitor isopropyl dodecylfluorophosphonate was employed for competitive activity-based protein profiling, leading to the identification of seven serine hydrolases with potential lipolytic activity. We demonstrated how a chemoproteomic approach can provide clues to the specificity of serine hydrolases by using a panel of neutral lipase inhibitors to identify an enzyme that reacts potently with a covalent monoacylglycerol lipase inhibitor. In combination with existing phenotypic data, our studies define a set of serine hydrolases that likely mediate critical metabolic reactions in asexual parasites and enable rational prioritization of future functional characterization and inhibitor development efforts.
- Published
- 2019
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