Sortase A mediated site-specific immobilization for identification of protein interactions in affinity purification-mass spectrometry experiments.
Proteomics
; 15(7): 1230-4, 2015 Apr.
Article
em En
| MEDLINE
| ID: mdl-25504886
ABSTRACT
Proteomics approaches using MS in combination with affinity purification have emerged as powerful tools to study protein-protein interactions. Here we make use of the specificity of sortase A transpeptidation reaction to prepare affinity matrices in which a protein bait is covalently linked to the matrix via a short C-terminal linker region. As a result of this site-directed immobilization, the bait remains functionally accessible to protein interactions. To apply this approach, we performed SILAC-based pull-down experiments and demonstrate the suitability of the approach.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Temas:
Geral
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
/
Cisteína Endopeptidases
/
Aminoaciltransferases
/
Mapeamento de Interação de Proteínas
/
Enzimas Imobilizadas
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Revista:
Proteomics
Assunto da revista:
BIOQUIMICA
Ano de publicação:
2015
Tipo de documento:
Article
País de afiliação:
Alemanha