%0 Journal Article
%T High-Throughput Screening for LC3/GABARAP Binders Utilizing the Fluorescence Polarization Assay.
%A Schwalm MP
%A Dopfer J
%A Knapp S
%A Rogov VV
%J Methods Mol Biol
%V 2845
%N 0
%D 2024
%M 39115669
暂无%R 10.1007/978-1-0716-4067-8_17
%X The characterization of interactions between autophagy modifiers (Atg8-family proteins) and their natural ligands (peptides and proteins) or small molecules is important for a detailed understanding of selective autophagy mechanisms and for the design of potential Atg8 inhibitors that affect the autophagy processes in cells. The fluorescence polarization (FP) assay is a rapid, cost-effective, and robust method that provides affinity and selectivity information for small molecules and peptide ligands targeting human Atg8 proteins.This chapter introduces the basic principles of FP assays. In addition, a case study on peptide interaction with human Atg8 proteins (LC3/GABARAPs) is described. Finally, data analysis and quality control of FP assays are discussed for the proper calculation of Ki values for the measured compounds.