Global profiling of dynamic protein palmitoylation

Nat Methods. 2011 Nov 6;9(1):84-9. doi: 10.1038/nmeth.1769.


The reversible thioester linkage of palmitic acid on cysteines, known as protein S-palmitoylation, facilitates the membrane association and proper subcellular localization of proteins. Here we report the metabolic incorporation of the palmitic acid analog 17-octadecynoic acid (17-ODYA) in combination with stable-isotope labeling with amino acids in cell culture (SILAC) and pulse-chase methods to generate a global quantitative map of dynamic protein palmitoylation events in cells. We distinguished stably palmitoylated proteins from those that turn over rapidly. Treatment with a serine lipase-selective inhibitor identified a pool of dynamically palmitoylated proteins regulated by palmitoyl-protein thioesterases. This subset was enriched in oncoproteins and other proteins linked to aberrant cell growth, migration and cancer. Our method provides a straightforward way to characterize global palmitoylation dynamics in cells and confirms enzyme-mediated depalmitoylation as a critical regulatory mechanism for a specific subset of rapidly cycling palmitoylated proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cysteine / metabolism*
  • Fatty Acids, Unsaturated / metabolism*
  • Lipase / antagonists & inhibitors
  • Lipoylation* / drug effects
  • Mass Spectrometry
  • Mice
  • Organophosphonates / pharmacology
  • Palmitic Acid / metabolism*
  • Protein Processing, Post-Translational
  • Serine Endopeptidases / metabolism
  • Thiolester Hydrolases / metabolism


  • Fatty Acids, Unsaturated
  • Organophosphonates
  • Palmitic Acid
  • 17-octadecynoic acid
  • Lipase
  • Thiolester Hydrolases
  • palmitoyl-protein thioesterase
  • Serine Endopeptidases
  • Cysteine