Use of analogs and inhibitors to study the functional significance of protein palmitoylation Journal Article


Author: Resh, M. D.
Article Title: Use of analogs and inhibitors to study the functional significance of protein palmitoylation
Abstract: Covalent attachment of palmitate to proteins is a post-translational modification that exerts diverse effects on protein localization and function. The three key technical approaches required for an investigator to determine the role of palmitoylation of your favorite palmitoylated protein (YFPP) are methods to: (1) detect YFPP palmitoylation; (2) alter or inhibit palmitoylation of YFPP; (3) determine the functional significance of altered YFPP palmitoylation. Here, I describe experimental methods to address these three issues. Both radioactive (radiolabeling with [3H]palmitate or 125I-IC16 palmitate) and non-radioactive (chemical labeling and mass spectrometry) methods to detect palmitoylated proteins are presented. Next, techniques to inhibit protein palmitoylation are described. These include site specific mutagenesis, and treatment of cells with inhibitors of protein palmitoylation, including 2-bromopalmitate, cerulenin, and tunicamycin. Alternative methods to replace palmitate with other fatty acids are also presented. Finally, general approaches to determining the effect of altered palmitoylation status on YFPP association with membranes and lipid rafts, as well as signal transduction, are described. © 2006 Elsevier Inc. All rights reserved.
Keywords: signal transduction; mass spectrometry; proteins; animals; protein; iodine 125; mutagenesis, site-directed; fatty acid; mutagenesis; acylation; lipid rafts; palmitoylation; lipid raft; fatty acylation; acyltransferases; palmitic acid; polyunsaturated fatty acids; 2-bromopalmitate; cerulenin; palmitate; plasma membrane; tunicamycin; 2 bromopalmitic acid
Journal Title: Methods
Volume: 40
Issue: 2
ISSN: 1046-2023
Publisher: Academic Press Inc., Elsevier Science  
Date Published: 2006-10-01
Start Page: 191
End Page: 197
Language: English
DOI: 10.1016/j.ymeth.2006.04.013
PUBMED: 17012032
PROVIDER: scopus
PMCID: PMC1712572
DOI/URL:
Notes: --- - "Cited By (since 1996): 52" - "Export Date: 4 June 2012" - "CODEN: MTHDE" - "Source: Scopus"
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  1. Marilyn D Resh
    103 Resh