Evaluation of prenylated peptides for use in cellular imaging and biochemical analysis

Joshua D. Ochocki, Urule Igbavboa, W. Gibson Wood, Edgar Arriaga, Elizabeth V Wattenberg, Mark D Distefano

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

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.

Original languageEnglish (US)
Title of host publicationTherapeutic Peptides
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages213-223
Number of pages11
ISBN (Print)9781627036726
DOIs
StatePublished - Jan 1 2014

Publication series

NameMethods in Molecular Biology
Volume1088
ISSN (Print)1064-3745

Fingerprint

Protein Prenylation
Prenylation
Peptides
Dimethylallyltranstransferase
ras Proteins
Terpenes
Capillary Electrophoresis
Enzyme Inhibitors
Confocal Microscopy
Flow Cytometry
Membrane Proteins
Research Personnel
Neoplasms
Proteins
Therapeutics

Keywords

  • Cell-penetrating peptide
  • Farnesyl
  • Lipid modification
  • Peptide
  • Posttranslational modification
  • Prenylation

Cite this

Ochocki, J. D., Igbavboa, U., Wood, W. G., Arriaga, E., Wattenberg, E. V., & Distefano, M. D. (2014). Evaluation of prenylated peptides for use in cellular imaging and biochemical analysis. In Therapeutic Peptides: Methods and Protocols (pp. 213-223). (Methods in Molecular Biology; Vol. 1088). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-673-3_14

Evaluation of prenylated peptides for use in cellular imaging and biochemical analysis. / Ochocki, Joshua D.; Igbavboa, Urule; Wood, W. Gibson; Arriaga, Edgar; Wattenberg, Elizabeth V; Distefano, Mark D.

Therapeutic Peptides: Methods and Protocols. Humana Press Inc., 2014. p. 213-223 (Methods in Molecular Biology; Vol. 1088).

Research output: Chapter in Book/Report/Conference proceedingChapter

Ochocki, JD, Igbavboa, U, Wood, WG, Arriaga, E, Wattenberg, EV & Distefano, MD 2014, Evaluation of prenylated peptides for use in cellular imaging and biochemical analysis. in Therapeutic Peptides: Methods and Protocols. Methods in Molecular Biology, vol. 1088, Humana Press Inc., pp. 213-223. https://doi.org/10.1007/978-1-62703-673-3_14
Ochocki JD, Igbavboa U, Wood WG, Arriaga E, Wattenberg EV, Distefano MD. Evaluation of prenylated peptides for use in cellular imaging and biochemical analysis. In Therapeutic Peptides: Methods and Protocols. Humana Press Inc. 2014. p. 213-223. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-673-3_14
Ochocki, Joshua D. ; Igbavboa, Urule ; Wood, W. Gibson ; Arriaga, Edgar ; Wattenberg, Elizabeth V ; Distefano, Mark D. / Evaluation of prenylated peptides for use in cellular imaging and biochemical analysis. Therapeutic Peptides: Methods and Protocols. Humana Press Inc., 2014. pp. 213-223 (Methods in Molecular Biology).
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