title: Evaluation of prenylated peptides for use in cellular imaging and biochemical analysis.
abstract: Protein prenylation involves the addition of a farnesyl (C15) or geranylgeranyl (C20) isoprenoid moiety onto the C-terminus of approximately 2 % of all mammalian proteins. This hydrophobic modification serves to direct membrane association of the protein. Due to the finding that the oncogenic protein Ras is naturally prenylated, several researchers have developed inhibitors of the prenyltransferase enzymes as cancer therapeutics. Despite numerous studies on the enzymology of prenylation in vitro, many questions remain about the process of prenylation in living cells. Using a combination of flow cytometry and confocal microscopy, we have shown that synthetic fluorescently labeled prenylated peptides enter a variety of different cell types. Additionally, using capillary electrophoresis we have shown that these peptides can be detected in minute quantities from lysates of cells treated with these peptides. This method will allow for further study of the enzymology of protein prenylation in living cells.
authors.affiliation: Department of Chemistry, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Ochocki
authors.forename: Urule
authors.initials: U
authors.lastname: Igbavboa
authors.forename: W Gibson
authors.initials: WG
authors.lastname: Wood
authors.forename: Edgar A
authors.initials: EA
authors.lastname: Arriaga
authors.forename: Elizabeth V
authors.initials: EV
authors.lastname: Wattenberg
authors.forename: Mark D
authors.initials: MD
authors.lastname: Distefano
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D002469:Cell Separation; D020374:Chromatography, Micellar Electrokinetic Capillary; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D057054:Molecular Imaging / Q000379:methods*; D009474:Neurons / Q000166:cytology; D010455:Peptides / Q000737:chemistry; D017368:Protein Prenylation; D051381:Rats
chemical_list: D005456:Fluorescent Dyes; D010455:Peptides