Cell Surface Biotinylation Using Furan Cross-Linking Chemistry.
Methods Mol Biol
; 2718: 11-21, 2023.
Article
em En
| MEDLINE
| ID: mdl-37665452
ABSTRACT
A detailed study of the cellular surfaceome poses major challenges for mass spectrometry analysis. Surface proteins are low abundant compared to intracellular proteins, and their inefficient extraction in aqueous medium leads to their aggregation and precipitation. To tackle such problems, surface biotinylation is frequently used to tag surface proteins with biotin, allowing for their enrichment, leading to a more sensitive mapping of surface proteomes. We here detail a new surface biotinylation protocol based on furan-biotin affinity purification to enrich plasma membrane proteins for proteomics. This protocol involves biotinylation of cell surface membrane proteins on viable cells, followed by affinity enrichment using streptavidin beads, trypsin digestion, peptide cleanup, and LC-MS/MS analysis.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Biotina
/
Espectrometria de Massas em Tandem
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
Bélgica