Site-specific analysis of protein S-acylation by resin-assisted capture.
J Lipid Res
; 52(2): 393-8, 2011 Feb.
Article
em En
| MEDLINE
| ID: mdl-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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas
/
Processamento de Proteína Pós-Traducional
/
Cisteína
/
Lipoilação
Idioma:
En
Revista:
J Lipid Res
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Estados Unidos