采用等温滴定量热法对 FOXO3-14-3-3 相互作用进行表征。

Characterization of FOXO3-14-3-3 Interaction by Isothermal Titration Calorimetry.

机构信息

Centre for Chemical Biology and Therapeutics, Institute for Stem Cell Science and Regenerative Medicine and National Centre for Biological Sciences-TIFR, Bangalore, India.

出版信息

Methods Mol Biol. 2025;2871:107-114. doi: 10.1007/978-1-0716-4217-7_10.

DOI:10.1007/978-1-0716-4217-7_10

PMID:39565582

Abstract

Isothermal titration calorimetry (ITC) is a well-established and convenient label-free technique for measuring biomolecular interactions in aqueous solutions. ITC enables accurate measurement of the affinities and thermodynamic parameters, such as changes in Gibbs energy, enthalpy, and entropy, which help dissect the binding mechanisms. ITC is commonly used to study protein-protein, protein-peptide, protein-DNA, and small-molecule inhibitor binding to target proteins. ITC can determine binding affinities ranging from nM to low-mM. This chapter provides a detailed protocol to investigate the binding of FOXO3 peptide to 14-3-3ε using ITC.

摘要

等温滴定量热法（ITC）是一种成熟且方便的无标记技术，可用于测量水溶液中的生物分子相互作用。ITC 能够准确测量亲和力和热力学参数，如吉布斯自由能、焓和熵的变化，这有助于剖析结合机制。ITC 常用于研究蛋白质-蛋白质、蛋白质-肽、蛋白质-DNA 和小分子抑制剂与靶蛋白的结合。ITC 可测定从纳摩尔到低毫摩尔范围的结合亲和力。本章提供了一个详细的方案，使用 ITC 研究 FOXO3 肽与 14-3-3ε 的结合。