2. Academic Validation
  3. Activity-Based Protein Profiling Identifies α-Ketoamides as Inhibitors for Phospholipase A2 Group XVI

Activity-Based Protein Profiling Identifies α-Ketoamides as Inhibitors for Phospholipase A2 Group XVI

  • ACS Chem Biol. 2019 Feb 15;14(2):164-169. doi: 10.1021/acschembio.8b00969.
Juan Zhou 1,  Elliot D Mock 1,  Andrea Martella 1,  Vasudev Kantae 1 2,  Xinyu Di 2,  Lindsey Burggraaff 3,  Marc P Baggelaar 1,  Karol Al-Ayed 1,  Alexander Bakker 1,  Bogdan I Florea 4,  Sebastian H Grimm 1,  Hans den Dulk 1,  Chun T Li 1,  Laura Mulder 1,  Herman S Overkleeft 4,  Thomas Hankemeier 2,  Gerard J P van Westen 3,  Mario van der Stelt 1
Affiliations

Affiliations

  • 1 Department of Molecular Physiology, Leiden Institute of Chemistry , Leiden University , Leiden , The Netherlands.
  • 2 Department of Analytical BioSciences and Metabolomics, Leiden Academic Centre for Drug Research , Leiden University , Leiden , The Netherlands.
  • 3 Department of Computational Drug Discovery, Leiden Academic Centre for Drug Research , Leiden University , Leiden , The Netherlands.
  • 4 Department of Bio-Organic Synthesis, Leiden Institute of Chemistry , Leiden University , Leiden , The Netherlands.
PMID: 30620559 DOI: 10.1021/acschembio.8b00969
Abstract

Phospholipase A2, group XVI (PLA2G16) is a thiol hydrolase from the HRASLS family that regulates lipolysis in adipose tissue and has been identified as a host factor enabling the cellular entry of picornaviruses. Chemical tools are essential to visualize and control PLA2G16 activity, but they have not been reported to date. Here, we show that MB064, which is a fluorescent Lipase probe, also labels recombinant and endogenously expressed PLA2G16. Competitive activity-based protein profiling (ABPP) using MB064 enabled the discovery of α-ketoamides as the first selective PLA2G16 inhibitors. LEI110 was identified as a potent PLA2G16 inhibitor ( Ki = 20 nM) that reduces cellular arachidonic acid levels and oleic acid-induced lipolysis in human HepG2 cells. Gel-based ABPP and chemical proteomics showed that LEI110 is a selective pan-inhibitor of the HRASLS family of thiol hydrolases (i.e., PLA2G16, HRASLS2, RARRES3 and iNAT). Molecular dynamic simulations of LEI110 in the reported crystal structure of PLA2G16 provided insight in the potential ligand-protein interactions to explain its binding mode. In conclusion, we have developed the first selective inhibitor that can be used to study the cellular role of PLA2G16.

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