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Methods Collections

Methods to study protein S-palmitoylation

Collection Overview

S-palmitoylation is the covalent attachment of the fatty acid palmitate to cysteine (Cys) residues of the substrate protein through thioester linkage. This universal post-translational lipid modification facilitates membrane association and therefore alters subcellular localization, stability and function of hundreds of proteins in cells. Among other lipid modifications of proteins, palmitoylation is unique because it is reversible and therefore can be regulated dynamically to influence various physiological processes within a cell. Palmitoylation occurs on diverse substrates, both soluble and transmembrane, and in a variety of different cell types and tissues and has crucial regulatory functions. Accumulating evidence indicates that malfunctions in palmitoylation are associated with cancer as well as a wide range of brain abnormalities, ranging from mild mental retardation to neurological disorders. Since its discovery 40 years ago, palmitoylation has been traditionally studied using metabolic radiolabeling that precluded the detection of dynamic as well as rare palmitoylation events. Over the last decade, a variety of different methods have been developed to study protein palmitoylation that overcome such limitations. Such methods broadly include chemical reporter-based metabolic labeling such as 17-octadecynoic acid (17-ODYA) labeling and biochemical assays such as and Acyl-PEG Exchange (APE), Acyl Biotin Exchange (ABE), and Acyl Resin-Assisted Capture (Acyl-RAC). Moreover, coupling these methods with other protein detection strategies has further expanded their applications. In this collection, we present the state-of-the-art techniques used to detect and enrich palmitoylated proteins that have opened up several exciting avenues for future research on dynamic palmitoylation.


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