Abstract
The endocannabinoid 2-arachidonoylglycerol (2-AG) is biosynthesized by diacylglycerol lipases DAGLα and DAGLβ. Chemical probes to perturb DAGLs are needed to characterize endocannabinoid function in biological processes. Here we report a series of 1,2,3-triazole urea inhibitors, along with paired negative-control and activity-based probes, for the functional analysis of DAGLβ in living systems. Optimized inhibitors showed high selectivity for DAGLβ over other serine hydrolases, including DAGLα (∼60-fold selectivity), and the limited off-targets, such as ABHD6, were also inhibited by the negative-control probe. Using these agents and Daglb(-/-) mice, we show that DAGLβ inactivation lowers 2-AG, as well as arachidonic acid and eicosanoids, in mouse peritoneal macrophages in a manner that is distinct and complementary to disruption of cytosolic phospholipase-A2. We observed a corresponding reduction in lipopolysaccharide-induced tumor necrosis factor-α release. These findings indicate that DAGLβ is a key metabolic hub within a lipid network that regulates proinflammatory responses in macrophages.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Animals
-
Arachidonic Acid / metabolism
-
Arachidonic Acids / biosynthesis
-
Cell Line
-
Cytokines / metabolism
-
Drug Discovery
-
Endocannabinoids / biosynthesis
-
Glycerides / biosynthesis
-
Inflammation / metabolism*
-
Lipid Metabolism / drug effects*
-
Lipoprotein Lipase / antagonists & inhibitors*
-
Lipoprotein Lipase / genetics
-
Lipoprotein Lipase / physiology
-
Macrophages, Peritoneal / drug effects*
-
Macrophages, Peritoneal / metabolism*
-
Mice
-
Mice, Knockout
-
Neurons / drug effects
-
Neurons / metabolism
-
Prostaglandins / metabolism
-
Protein Isoforms
-
Proteome / drug effects
-
Quantitative Structure-Activity Relationship
-
Signal Transduction / drug effects
-
Triazoles / chemical synthesis
-
Triazoles / pharmacology
-
Tumor Necrosis Factor-alpha / metabolism
Substances
-
Arachidonic Acids
-
Cytokines
-
Endocannabinoids
-
Glycerides
-
Prostaglandins
-
Protein Isoforms
-
Proteome
-
Triazoles
-
Tumor Necrosis Factor-alpha
-
Arachidonic Acid
-
glyceryl 2-arachidonate
-
Lipoprotein Lipase
-
diacylglycerol lipase beta, mouse
Associated data
-
PubChem-Substance/144223306
-
PubChem-Substance/144223307
-
PubChem-Substance/144223308
-
PubChem-Substance/144223309
-
PubChem-Substance/144223310
-
PubChem-Substance/144223311
-
PubChem-Substance/144223312
-
PubChem-Substance/144223313
-
PubChem-Substance/144223314
-
PubChem-Substance/144223315
-
PubChem-Substance/144223316
-
PubChem-Substance/144223317
-
PubChem-Substance/144223318
-
PubChem-Substance/144223319
-
PubChem-Substance/144223320
-
PubChem-Substance/144223321
-
PubChem-Substance/144223322
-
PubChem-Substance/144223323
-
PubChem-Substance/144223324
-
PubChem-Substance/144223325
-
PubChem-Substance/144223326
-
PubChem-Substance/144223327
-
PubChem-Substance/144223328
-
PubChem-Substance/144223329
-
PubChem-Substance/144223330
-
PubChem-Substance/144223331
-
PubChem-Substance/144223332
-
PubChem-Substance/144223333
-
PubChem-Substance/144223334
-
PubChem-Substance/144223335
-
PubChem-Substance/144223336
-
PubChem-Substance/144223337
-
PubChem-Substance/144223338
-
PubChem-Substance/144223339
-
PubChem-Substance/144223340
-
PubChem-Substance/144223341
-
PubChem-Substance/144223342
-
PubChem-Substance/144223343
-
PubChem-Substance/144223344