An improved protein lipid overlay assay for studying lipid-protein interactions.

ABSTRACT:

Background

Lipids perform multiple functions in the cell, and lipid-protein interactions play a key role in metabolism. Although various techniques have been developed to study lipid-protein interactions, the interacting protein partners that bind to most lipids remain unknown. The protein lipid overlay (PLO) assay has revealed numerous lipid-protein interactions, but its application suffers from unresolved technical issues.

Results

Herein, we found that blocking proteins may interfere with interactions between lipids and their binding proteins if a separate blocking step is carried out before the incubation step in the PLO assay. To overcome this, we modified the PLO assay by combining an incubation step alongside the blocking step. Verification experiments included phospha

SUBMITTER: Han X 

PROVIDER: S-EPMC7060618 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature