Book chapter
A Fluorescence-Based Assay to Determine PDZ-Ligand Binding Thermodynamics: Methods and Protocols
PDZ Mediated Interactions, pp.137-148
Methods in Molecular Biology, v. 2256, Humana Press
2021
DOI: 10.1007/978-1-0716-1166-1_8
PMCID: PMC8507397
PMID: 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.
Details
- Title: Subtitle
- A Fluorescence-Based Assay to Determine PDZ-Ligand Binding Thermodynamics: Methods and Protocols
- Creators
- Young Joo Sun - University of IowaErnesto J Fuentes - Univ Iowa, Dept Biochem, Iowa City, IA 52242 USA
- Contributors
- J P Borg (Editor)
- Resource Type
- Book chapter
- Publication Details
- PDZ Mediated Interactions, pp.137-148
- Series
- Methods in Molecular Biology; v. 2256
- DOI
- 10.1007/978-1-0716-1166-1_8
- PMID
- 34014520
- PMCID
- PMC8507397
- NLM abbreviation
- Methods Mol Biol
- eISSN
- 1940-6029
- ISSN
- 1064-3745
- Publisher
- Humana Press; New York
- Number of pages
- 12
- Grant note
- R21-AI135305 / National Institutes of Health; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA
- Language
- English
- Date published
- 2021
- Academic Unit
- Biochemistry and Molecular Biology
- Record Identifier
- 9984293083902771
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