Identification of the interacting partners of a lysosomal membrane protein in living cells by BioID technique.
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ABSTRACT:
SUBMITTER: Nguyen-Tien D
PROVIDER: S-EPMC8983420 | biostudies-literature | 2022 Jun
REPOSITORIES: biostudies-literature
Publications
Identification of the interacting partners of a lysosomal membrane protein in living cells by BioID technique.
Nguyen-Tien Dat D Suzuki Takehiro T Kobayashi Toshihiko T Toyama-Sorimachi Noriko N Dohmae Naoshi N
STAR protocols 20220331 2
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 m ...[more]
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