利用圆二色性测定蛋白质折叠与变性剂、渗透压调节剂或配体的关系。
Determination of the folding of proteins as a function of denaturants, osmolytes or ligands using circular dichroism.
作者信息
Greenfield Norma J
机构信息
Department of Neuroscience and Cell Biology, Robert Wood Johnson Medical School, 675 Hoes Lane West, Piscataway, New Jersey 08854-8021, USA.
出版信息
Nat Protoc. 2006;1(6):2733-41. doi: 10.1038/nprot.2006.229.
Abstract
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.
摘要
圆二色性（CD）是研究蛋白质相互作用和稳定性的一种出色工具。本方案涵盖了获取和分析圆二色光谱的方法，以测量蛋白质折叠随变性剂、渗透压调节剂或配体的变化。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。这些实验需要2至5小时。
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本文引用的文献
1
Computer calculation of equilibrium concentrations in mixtures of metal ions and complexing species.
Talanta. 1967 Jul;14(7):833-42. doi: 10.1016/0039-9140(67)80105-x.
2
Using circular dichroism spectra to estimate protein secondary structure.
Nat Protoc. 2006;1(6):2876-90. doi: 10.1038/nprot.2006.202.
3
Using circular dichroism collected as a function of temperature to determine the thermodynamics of protein unfolding and binding interactions.
Nat Protoc. 2006;1(6):2527-35. doi: 10.1038/nprot.2006.204.
4
Probing protein binding sites by circular dichroism spectroscopy.
Curr Drug Discov Technol. 2004 Jun;1(2):133-53. doi: 10.2174/1570163043335135.
5
Osmolyte-induced protein folding free energy changes.
Proteins. 2006 May 1;63(2):290-6. doi: 10.1002/prot.20868.
6
Spectroscopic identification of binding modes of anthracene probes and DNA sequence recognition.
Photochem Photobiol. 2006 Jan-Feb;82(1):20-30. doi: 10.1562/2005-05-24-RA-539.
7
Thermodynamics of denaturant-induced unfolding of a protein that exhibits variable two-state denaturation.
Biochemistry. 2004 Oct 26;43(42):13357-69. doi: 10.1021/bi048666j.
8
Circular dichroism analysis for protein-protein interactions.
Methods Mol Biol. 2004;261:55-78. doi: 10.1385/1-59259-762-9:055.
9
Analysis of circular dichroism data.
Methods Enzymol. 2004;383:282-317. doi: 10.1016/S0076-6879(04)83012-X.
10
Tropomodulin contains two actin filament pointed end-capping domains.
J Biol Chem. 2003 Oct 10;278(41):40000-9. doi: 10.1074/jbc.M306895200. Epub 2003 Jul 14.
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