Identification of the major prostaglandin glycerol ester hydrolase in human cancer cells

J Biol Chem. 2014 Dec 5;289(49):33741-53. doi: 10.1074/jbc.M114.582353. Epub 2014 Oct 9.

Abstract

Prostaglandin glycerol esters (PG-Gs) are produced as a result of the oxygenation of the endocannabinoid, 2-arachidonoylglycerol, by cyclooxygenase 2. Understanding the role that PG-Gs play in a biological setting has been difficult because of their sensitivity to enzymatic hydrolysis. By comparing PG-G hydrolysis across human cancer cell lines to serine hydrolase activities determined by activity-based protein profiling, we identified lysophospholipase A2 (LYPLA2) as a major enzyme responsible for PG-G hydrolysis. The principal role played by LYPLA2 in PGE2-G hydrolysis was confirmed by siRNA knockdown. Purified recombinant LYPLA2 hydrolyzed PG-Gs in the following order of activity: PGE2-G > PGF2α-G > PGD2-G; LYPLA2 hydrolyzed 1- but not 2-arachidonoylglycerol or arachidonoylethanolamide. Chemical inhibition of LYPLA2 in the mouse macrophage-like cell line, RAW264.7, elicited an increase in PG-G production. Our data indicate that LYPLA2 serves as a major PG-G hydrolase in human cells. Perturbation of this enzyme should enable selective modulation of PG-Gs without alterations in endocannabinoids, thereby providing a means to decipher the unique functions of PG-Gs in biology and disease.

Keywords: Activity-based Protein Profiling; Cancer; Chemical Biology; Endocannabinoids; Enzyme Kinetics; Lipid Metabolism; Prostaglandin; Prostaglandin Glycerol Esters; Serine Hydrolase.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Arachidonic Acids / metabolism
  • Cell Line, Tumor
  • Cyclooxygenase 2 / genetics
  • Cyclooxygenase 2 / metabolism
  • Endocannabinoids / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Esters
  • Gene Expression Regulation, Neoplastic*
  • Glycerides / metabolism
  • Glycerol / metabolism*
  • Humans
  • Hydrolysis
  • Kinetics
  • Macrophages / cytology
  • Macrophages / enzymology*
  • Mice
  • Polyunsaturated Alkamides / metabolism
  • Prostaglandins / chemistry
  • Prostaglandins / metabolism*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Signal Transduction
  • Substrate Specificity
  • Thiolester Hydrolases / antagonists & inhibitors
  • Thiolester Hydrolases / genetics
  • Thiolester Hydrolases / metabolism*

Substances

  • Arachidonic Acids
  • Endocannabinoids
  • Esters
  • Glycerides
  • Polyunsaturated Alkamides
  • Prostaglandins
  • RNA, Small Interfering
  • Recombinant Proteins
  • glyceryl 2-arachidonate
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • LYPLA2 protein, human
  • Thiolester Hydrolases
  • Glycerol
  • anandamide