A proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells.
STAR Protoc
; 2(4): 100962, 2021 12 17.
Article
em En
| MEDLINE
| ID: mdl-34820639
ABSTRACT
Some protein-protein interactions are induced by different kinds of stimulation and are dependent on specific genes. To identify these interaction partners, we present a protocol which utilizes affinity purification of Flag-tagged protein complexes followed by mass-spectrometry-based proteomics to compare stimulation-induced interactomes between wild-type and CRISPR-Cas9-mediated knockout cells. The candidates of interest are identified using bioinformatic analyses and verified by biochemical approaches. This protocol is highly versatile and applies to a variety of cells and different types of stimulation. For complete details on the use and execution of this protocol, please refer to (Zhu et al., 2021).
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Espectrometria de Massas
/
Proteínas
/
Mapeamento de Interação de Proteínas
/
Proteômica
Limite:
Humans
Idioma:
En
Revista:
STAR Protoc
Ano de publicação:
2021
Tipo de documento:
Article
País de afiliação:
China