Isolation and determination of PI(4,5)P2 proximal proteins

ABSTRACT: We screened proteins in close proximity to PI(4,5)P2 in epithelial cells. We established HaCaT cells stably expressing APEX2 N-terminally fused to the PH domain of PLCd1, which specifically binds to PI(4,5)P2. In the presence of hydrogen peroxide and biotin-phenol, proteins proximal to APEX2 are biotinylated by APEX2, allowing for their enrichment using streptavidin beads. APEX2-fused non-PI(4,5) P2-binding mutant of the PH domain of PLCd1 (R40L) was used as the negative control.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Yoshikazu Nakamura  

LAB HEAD: Yoshikazu nakamura

PROVIDER: PXD031266 | Pride | 2022-03-26

REPOSITORIES: Pride