Site-specific analysis of protein S-acylation by resin-assisted capture.
Published
Journal Article
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.
Full Text
Duke Authors
Cited Authors
- Forrester, MT; Hess, DT; Thompson, JW; Hultman, R; Moseley, MA; Stamler, JS; Casey, PJ
Published Date
- February 2011
Published In
Volume / Issue
- 52 / 2
Start / End Page
- 393 - 398
PubMed ID
- 21044946
Pubmed Central ID
- 21044946
Electronic International Standard Serial Number (EISSN)
- 1539-7262
Digital Object Identifier (DOI)
- 10.1194/jlr.D011106
Language
- eng
Conference Location
- United States