Cellular palmitoylation and trafficking of lipidated peptides

Jeremiah M. Draper, Zuping Xia, Charles D. Smith

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Many important signaling proteins require the posttranslational addition of fatty acid chains for their proper subcellular localization and function. One such modification is the addition of palmitoyl moieties by enzymes known as palmitoyl acyltransferases (PATs). Substrates for PATs include C-terminally farnesylated proteins, such as H- and N-Ras, as well as N-terminally myristoylated proteins, such as many Src-related tyrosine kinases. The molecular and biochemical characterization of PATs has been hindered by difficulties in developing effective methods for the analysis of PAT activity. In this study, we describe the use of cell-permeable, fluorescently labeled lipidated peptides that mimic the PAT recognition domains of farnesylated and myristoylated proteins. These PAT substrate mimetics are accumulated by SKOV3 cells in a saturable and time-dependent manner. Although both peptides are rapidly palmitoylated, the SKOV3 cells have a greater capacity to palmitoylate the myristoylated peptide than the farnesylated peptide. Confocal microscopy indicated that the palmitoylated peptides colocalized with Golgi and plasma membrane markers, whereas the corresponding nonpalmitoylatable peptides accumulated in the Golgi but did not traffic to the plasma membrane. Overall, these studies indicate that the lipidated peptides provide useful cellular probes for quantitative and compartmentalization studies of protein palmitoylation in intact cells.

Original languageEnglish (US)
Pages (from-to)1873-1884
Number of pages12
JournalJournal of Lipid Research
Volume48
Issue number8
DOIs
StatePublished - Aug 1 2007

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Lipoylation
Acyltransferases
Peptides
Cell membranes
Proteins
Cell Membrane
src-Family Kinases
Confocal microscopy
Substrates
Confocal Microscopy
Protein-Tyrosine Kinases
Fatty Acids
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

Draper, Jeremiah M. ; Xia, Zuping ; Smith, Charles D. / Cellular palmitoylation and trafficking of lipidated peptides. In: Journal of Lipid Research. 2007 ; Vol. 48, No. 8. pp. 1873-1884.
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Cellular palmitoylation and trafficking of lipidated peptides. / Draper, Jeremiah M.; Xia, Zuping; Smith, Charles D.

In: Journal of Lipid Research, Vol. 48, No. 8, 01.08.2007, p. 1873-1884.

Research output: Contribution to journalArticle

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