A Fluorescence-Based Assay to Determine PDZ-Ligand Binding Thermodynamics.
Methods Mol Biol
; 2256: 137-148, 2021.
Article
em En
| MEDLINE
| ID: mdl-34014520
ABSTRACT
Postsynaptic density-95, disks-large, and zonula occludens-1 (PDZ) domain interactions with cognate linear binding motifs (i.e., PDZ-binding motifs or PBMs) are important for many biological processes and can be pathological when disrupted. There are hundreds of PDZ-PBM interactions reported but few have been quantitatively determined. Moreover, PDZ-PBM interactions have been identified as potential therapeutic targets. To thoroughly understand PDZ-PBM binding energetics and their specificity, we have developed a sensitive and quantitative equilibrium binding assay. Here, we describe a protocol for determining PDZ-PBM binding energetics using fluorescence anisotropy-based methodology.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Termodinâmica
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Proteínas Supressoras de Tumor
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Guanilato Quinases
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Domínios PDZ
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Fluorescência
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Polarização de Fluorescência
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Proteínas de Membrana
Tipo de estudo:
Guideline
Limite:
Humans
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article