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Defining Substrate Specificities for Lipase and Phospholipase Candidates
Published on: November 23, 2016
Activity-based probes that target functional subclasses of phospholipases in proteomes
Sarah E Tully1, Benjamin F Cravatt
1Department of Chemical Physiology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|February 25, 2010
Summary
New activity-based protein profiling (ABPP) probes target phospholipases, enzymes crucial for cell membranes and lipid signaling. These chemical tools reveal enzyme substrate specificities, aiding in the discovery of novel lipid-metabolizing enzymes.
Area of Science:
- Biochemistry
- Enzymology
- Chemical Biology
Background:
- Phospholipases are critical enzymes metabolizing cell membrane phospholipids and regulating lipid-signaling pathways.
- Discovering novel phospholipases requires selective chemical probes that identify enzymes based on substrate specificity and catalytic activity.
Purpose of the Study:
- To synthesize and characterize novel activity-based protein profiling (ABPP) probes targeting phospholipases within the serine hydrolase superfamily.
- To demonstrate the utility of these ABPP probes in identifying phospholipase substrate specificities in complex biological systems.
Main Methods:
- Synthesis of ABPP probes incorporating recognition and reactivity elements specific to phospholipases.
- Application of ABPP probes to cell and tissue proteomes to profile phospholipase activity.
- Characterization of probe interactions to determine sn-1 and sn-2 substrate specificities.
Main Results:
- Developed and validated ABPP probes for targeting serine hydrolase superfamily phospholipases.
- Demonstrated accurate reporting of sn-1 and sn-2 substrate specificities for phospholipases in proteomic samples.
- Identified the sn-1-selective phospholipase DDHD1 and a calcium-dependent transacylase involved in endocannabinoid biosynthesis.
Conclusions:
- Phospholipase-directed ABPP probes are effective tools for discovering and characterizing novel lipid-metabolizing enzymes.
- These probes provide valuable insights into the substrate preferences of phospholipases.
- The developed ABPP strategy facilitates advancements in understanding lipid metabolism and signaling pathways.
