利用圆二色性测定蛋白质折叠与变性剂、渗透剂或配体的关系

IF 13.1 1区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Nature Protocols Pub Date : 2007-01-11 DOI:10.1038/nprot.2006.229

Norma J Greenfield

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引用次数: 135

摘要

圆二色性（CD）是研究蛋白质相互作用和稳定性的绝佳工具。本程序涵盖了获取和分析圆二色光谱的方法，用于测量蛋白质折叠随变性剂、渗透溶质或配体的变化而发生的变化。其应用包括测定蛋白质折叠的自由能、突变对蛋白质稳定性的影响以及估算蛋白质与其他蛋白质、DNA 或配体（如底物或抑制剂）相互作用的结合常数。实验需要 2-5 小时。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Determination of the folding of proteins as a function of denaturants, osmolytes or ligands using circular dichroism

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Determination of the folding of proteins as a function of denaturants, osmolytes or ligands using circular dichroism

Circular dichroism (CD) is an excellent tool for examining the interactions and stability of proteins. This protocol covers methods to obtain and analyze circular dichroism spectra to measure changes in the folding of proteins as a function of denaturants, osmolytes or ligands. Applications include determination of the free energy of folding of a protein, the effects of mutations on protein stability and the estimation of binding constants for the interactions of proteins with other proteins, DNA or ligands, such as substrates or inhibitors. The experiments require 2–5 h.

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来源期刊

Nature Protocols 生物-生化研究方法

CiteScore

29.10

自引率

0.70%

发文量

128

审稿时长

4 months

期刊介绍： Nature Protocols focuses on publishing protocols used to address significant biological and biomedical science research questions, including methods grounded in physics and chemistry with practical applications to biological problems. The journal caters to a primary audience of research scientists and, as such, exclusively publishes protocols with research applications. Protocols primarily aimed at influencing patient management and treatment decisions are not featured. The specific techniques covered encompass a wide range, including but not limited to: Biochemistry, Cell biology, Cell culture, Chemical modification, Computational biology, Developmental biology, Epigenomics, Genetic analysis, Genetic modification, Genomics, Imaging, Immunology, Isolation, purification, and separation, Lipidomics, Metabolomics, Microbiology, Model organisms, Nanotechnology, Neuroscience, Nucleic-acid-based molecular biology, Pharmacology, Plant biology, Protein analysis, Proteomics, Spectroscopy, Structural biology, Synthetic chemistry, Tissue culture, Toxicology, and Virology.