Related Experiment Video
Updated: Apr 9, 2026
10:50
Molecular Modulation by Lentivirus-Delivered Specific shRNAs in Endoplasmic Reticulum Stressed Neurons
Published on: April 24, 2021
1.4K
Highly Selective, Reversible Inhibitor Identified by Comparative Chemoproteomics Modulates Diacylglycerol Lipase
Marc P Baggelaar1, Pascal J P Chameau2, Vasudev Kantae3
1†Department of Bioorganic Synthesis, Leiden Institute of Chemistry, Leiden University, Leiden 2300 RA, The Netherlands.
Journal of the American Chemical Society
|June 18, 2015
Summary
Researchers developed a novel, selective inhibitor, LEI105, for diacylglycerol lipase (DAGL)-α and -β enzymes. This discovery aids in studying endocannabinoid signaling and provides evidence for 2-AG
Area of Science:
- Biochemistry
- Neuroscience
- Pharmacology
Background:
- Diacylglycerol lipase (DAGL)-α and -β are key enzymes in synthesizing 2-arachidonoylglycerol (2-AG), a crucial endocannabinoid.
- Selective and reversible inhibitors are needed to investigate DAGL function in neuronal cells.
- Current inhibitors lack the required selectivity and temporal control for acute studies.
Purpose of the Study:
- To identify and characterize a highly selective inhibitor for DAGL-α and DAGL-β.
- To utilize structure-guided design and chemoproteomics for inhibitor discovery and selectivity profiling.
- To investigate the role of 2-AG in retrograde signaling via inhibition of DAGL.
Main Methods:
- Employed a chemoproteomics strategy combining comparative and competitive activity-based protein profiling (ABPP).
- Utilized broad-spectrum fluorophosphonate and specific β-lactone probes to identify inhibitors in complex proteomes.
- Validated inhibitor selectivity against other endocannabinoid-metabolizing enzymes and cannabinoid receptors.
Main Results:
- Discovered LEI105, an α-ketoheterocycle, as a potent, selective, and reversible dual DAGL-α/DAGL-β inhibitor.
- LEI105 demonstrated no inhibition of related enzymes (ABHD6, ABHD12, MAGL, FAAH) or affinity for CB1 receptors.
- LEI105 reduced 2-AG levels in Neuro2A cells and inhibited CB1-receptor-mediated synaptic plasticity in mouse hippocampal slices.
Conclusions:
- LEI105 is a valuable pharmacological tool for studying DAGL function and endocannabinoid signaling.
- The findings support the hypothesis of "on demand" 2-AG biosynthesis in retrograde signaling.
- This study highlights the power of ABPP in discovering selective enzyme inhibitors within native biological systems.
Related Concept Videos
01:30Ligand-Gated Ion Channel Receptor: Gating Mechanism
4.9K
Ligand-gated ion channels are transmembrane proteins that play a vital role in intercellular communication and functions of the nervous system. They allow the influx of ions across the membrane once the neurotransmitter binds, allowing the subsequent transmission of electrical excitation across the neurons. Other ligand-gated ion channels, like the γ-aminobutyric acid (GABA) receptor, permit anions like chloride into the cells on the binding of the GABA molecule. Their entry into the cell...
4.9K
01:18Indirect-Acting Cholinergic Agonists: Mechanism of Action
3.0K
Indirect-acting cholinergic agonists work by interacting with an enzyme called acetylcholinesterase (AChE) in the synaptic cleft. They can be reversible or irreversible inhibitors and have different effects on the enzyme.
Reversible inhibitors like edrophonium bind to a specific part of the enzyme called the anionic catalytic site. They form noncovalent bonds, which means they are not strongly attached to the enzyme. This creates a temporary and less stable enzyme–inhibitor complex,...
Reversible inhibitors like edrophonium bind to a specific part of the enzyme called the anionic catalytic site. They form noncovalent bonds, which means they are not strongly attached to the enzyme. This creates a temporary and less stable enzyme–inhibitor complex,...
3.0K
01:30Enzyme Inhibition
95.2K
Inhibitors are molecules that reduce enzyme activity by binding to the enzyme. In a normally functioning cell, enzymes are regulated by a variety of inhibitors. Drugs and other toxins can also inhibit enzymes. Some inhibitors bind to the enzyme’s active site, while others inhibit enzymatic activity by binding to other sites on the protein structure.
95.2K
01:29Indirect-Acting Cholinergic Agonists: Chemistry and Structure-Activity Relationship
1.2K
Indirect-acting cholinergic agonists are agents that interact with the acetylcholinesterase enzyme in the synaptic cleft, preventing the breakdown of acetylcholine into choline and acetate. Consequently, the concentration of acetylcholine in the synaptic cleft increases. These agonists can be classified into reversible and irreversible inhibitors based on their duration of action.
Reversible inhibitors display short to medium durations of action. Short-acting agents include simple alcohols with...
Reversible inhibitors display short to medium durations of action. Short-acting agents include simple alcohols with...
1.2K
00:46Feedback Inhibition
58.8K
Biochemical reactions are occurring constantly in cells, converting starting substances to different products, usually with the help of enzymes that speed the reactions. Without enzymes, it would take far too long for most reactions to occur to be useful to the cell!
58.8K
01:11IP3/DAG Signaling Pathway
16.0K
Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and...
16.0K