Journal article
Site-specific analysis of protein S-acylation by resin-assisted capture[S]
Journal of lipid research, Vol.52(2), pp.393-398
02/2011
DOI: 10.1194/jlr.D011106
PMCID: PMC3023561
PMID: 21044946
Abstract
Protein S-acylation is a major posttranslational modification whereby a cysteine thiol is converted to a thioester. A prototype is S-palmitoylation (fatty acylation), in which a protein undergoes acylation with a hydrophobic 16 carbon lipid chain. Although this modification is a well-recognized determinant of protein function and localization, current techniques to study cellular S-acylation are cumbersome and/or technically demanding. We recently described a simple and robust methodology to rapidly identify S-nitrosylation sites in proteins via resin-assisted capture (RAC) and provided an initial description of the applicability of the technique to S-acylated proteins (acyl-RAC). Here we expand on the acyl-RAC assay, coupled with mass spectrometry-based proteomics, to characterize both previously reported and novel sites of endogenous S-acylation. Acyl-RAC should therefore find general applicability in studies of both global and individual protein S-acylation in mammalian cells.
Details
- Title: Subtitle
- Site-specific analysis of protein S-acylation by resin-assisted capture[S]
- Creators
- Michael T Forrester - Department of Biochemistry, Duke University Medical Center, Durham, NC 27710Douglas T Hess - Institute for Transformative Molecular Medicine, Case Western Reserve University and University Hospitals, Cleveland, OH 44106J. Will Thompson - Department of Proteomics Core Facility, Duke University Medical Center, Durham, NC 27710Rainbo Hultman - Department of Biochemistry, Duke University Medical Center, Durham, NC 27710M. Arthur Moseley - Department of Proteomics Core Facility, Duke University Medical Center, Durham, NC 27710Jonathan S Stamler - Institute for Transformative Molecular Medicine, Case Western Reserve University and University Hospitals, Cleveland, OH 44106Patrick J Casey - Department of Biochemistry, Duke University Medical Center, Durham, NC 27710
- Resource Type
- Journal article
- Publication Details
- Journal of lipid research, Vol.52(2), pp.393-398
- DOI
- 10.1194/jlr.D011106
- PMID
- 21044946
- PMCID
- PMC3023561
- NLM abbreviation
- J Lipid Res
- ISSN
- 0022-2275
- eISSN
- 1539-7262
- Publisher
- Elsevier Inc
- Language
- English
- Date published
- 02/2011
- Academic Unit
- Molecular Physiology and Biophysics; Psychiatry; Iowa Neuroscience Institute
- Record Identifier
- 9984065854102771
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