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Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange (ABE)
Published on: February 18, 2013
Global profiling of dynamic protein palmitoylation
Brent R Martin1, Chu Wang, Alexander Adibekian
1The Department of Chemical Physiology and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA.
Nature Methods
|November 8, 2011
Summary
Researchers mapped dynamic protein palmitoylation using a novel metabolic labeling method. This reveals key regulatory roles of depalmitoylation in cancer-related proteins.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Protein S-palmitoylation, the attachment of palmitic acid to cysteines via a thioester bond, is crucial for protein membrane association and localization.
- Understanding the dynamics of palmitoylation is essential for deciphering cellular signaling and protein function.
Purpose of the Study:
- To develop a method for globally mapping dynamic protein palmitoylation events in cells.
- To distinguish between stable and transient palmitoylation.
- To identify proteins regulated by dynamic palmitoylation and their functional implications.
Main Methods:
- Metabolic incorporation of 17-octadecynoic acid (17-ODYA), a palmitic acid analog.
- Stable-isotope labeling with amino acids in cell culture (SILAC) for quantitative analysis.
- Pulse-chase experiments to track palmitoylation turnover.
- Treatment with a serine lipase-selective inhibitor to probe depalmitoylation.
Main Results:
- Generated a global quantitative map of dynamic protein palmitoylation.
- Differentiated between stably and rapidly turning over palmitoylated proteins.
- Identified a subset of dynamically palmitoylated proteins regulated by palmitoyl-protein thioesterases.
- This subset showed enrichment in oncoproteins and proteins associated with cancer-related cellular processes.
Conclusions:
- The developed method enables straightforward characterization of global palmitoylation dynamics.
- Enzyme-mediated depalmitoylation is a critical regulatory mechanism for rapidly cycling palmitoylated proteins.
- Dynamic palmitoylation plays a significant role in regulating proteins involved in cell growth, migration, and cancer.
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