Activity-based protein profiling identifies α-ketoamides as inhibitors for Phospholipase A2 Group XVI

Author(s):
Zhou, J.; Mock, E.D. (0000-0002-1500-2856); Martella, A. (0000-0002-4680-7702); Kantae, V. (0000-0001-8541-0882); Di, X.; Burggraaff, L. (0000-0002-2442-0443); Baggelaar, M.P.; Al-Ayed, K.; Bakker, A.; Florea, B.I. (0000-0001-7114-2266); Grimm, S.H. (0000-0002-8832-8259); Dulk, H. den; Li, C.T.; Mulder, L.; Overkleeft, H.S. (0000-0001-6992-4760); Westen, G.J.P. van (0000-0003-0717-1817); Hankemeier, T. (0000-0001-7871-2073); Stelt, M. van der (0000-0002-1029-5717)
Published
2019
Publication type
Article / Letter to editor

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 have not been reported to date. Here we show that MB064, 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.

Date issued
2019-01-08
Language
English
Is part of
ACS Chemical Biology (2019), Vol. 14, no. 2, p. 164-169
hdl
1887/69904
doi
10.1021/acschembio.8b00969