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Autores principales: Schwalm, Martin P., Dopfer, Johannes, Knapp, Stefan, Rogov, Vladimir V.
Formato: Preprint
Publicado: 2024
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Acceso en línea:https://arxiv.org/abs/2401.04453
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author Schwalm, Martin P.
Dopfer, Johannes
Knapp, Stefan
Rogov, Vladimir V.
author_facet Schwalm, Martin P.
Dopfer, Johannes
Knapp, Stefan
Rogov, Vladimir V.
contents 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 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 which provides affinity and selectivity information of small molecules and peptide ligands targeting human Atg8 proteins. This chapter introduces the basic principles of FP assays are introduced. In addition, a case study for FP small molecule and peptide interactions with human Atg8 proteins (LC3/GABARAPs) is described. Finally, data analysis and quality control of FP studies are discussed for the proper calculation of Ki values of the measured compounds.
format Preprint
id arxiv_https___arxiv_org_abs_2401_04453
institution arXiv
publishDate 2024
record_format arxiv
spellingShingle High throughput screening for LC3/GABARAP binders utilizing fluorescence polarization assay
Schwalm, Martin P.
Dopfer, Johannes
Knapp, Stefan
Rogov, Vladimir V.
Biomolecules
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 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 which provides affinity and selectivity information of small molecules and peptide ligands targeting human Atg8 proteins. This chapter introduces the basic principles of FP assays are introduced. In addition, a case study for FP small molecule and peptide interactions with human Atg8 proteins (LC3/GABARAPs) is described. Finally, data analysis and quality control of FP studies are discussed for the proper calculation of Ki values of the measured compounds.
title High throughput screening for LC3/GABARAP binders utilizing fluorescence polarization assay
topic Biomolecules
url https://arxiv.org/abs/2401.04453