Identification of the interacting partners of a lysosomal membrane protein in living cells by BioID technique.
STAR Protoc
; 3(2): 101263, 2022 06 17.
Article
em En
| MEDLINE
| ID: mdl-35403001
ABSTRACT
The purpose of this protocol is to screen and identify the physiologically relevant interactors of a lysosomal protein in living cells. Here, we describe how to identify solute carrier family 15 member 4 (SLC15A4)-interacting proteins by BioID and mass spectrometry analysis. This protocol utilizes fusion of SLC15A4 with a mutant form of biotin ligase, BirA. The fusion protein can promiscuously biotinylate the proteins proximal to SLC15A4. The biotinylated endogenous proteins are pulled down by magnetic streptavidin beads and detected by mass spectrometry analysis. For complete details on the use and execution of this protocol, please refer to Kobayashi et al. (2021).
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article