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

International Standard Serial Number (ISSN)

  • 0022-2275

Digital Object Identifier (DOI)

  • 10.1194/jlr.D011106

Language

  • eng