利用圆二色光谱估计蛋白质二级结构。
Using circular dichroism spectra to estimate protein secondary structure.
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):2876-90. doi: 10.1038/nprot.2006.202.
Abstract
Circular dichroism (CD) is an excellent tool for rapid determination of the secondary structure and folding properties of proteins that have been obtained using recombinant techniques or purified from tissues. The most widely used applications of protein CD are to determine whether an expressed, purified protein is folded, or if a mutation affects its conformation or stability. In addition, it can be used to study protein interactions. This protocol details the basic steps of obtaining and interpreting CD data, and methods for analyzing spectra to estimate the secondary structural composition of proteins. CD has the advantage that measurements may be made on multiple samples containing < or =20 microg of proteins in physiological buffers in a few hours. However, it does not give the residue-specific information that can be obtained by x-ray crystallography or NMR.
摘要
圆二色性（CD）是一种出色的工具，可用于快速测定通过重组技术获得或从组织中纯化的蛋白质的二级结构和折叠特性。蛋白质CD最广泛的应用是确定表达并纯化的蛋白质是否折叠，或者突变是否影响其构象或稳定性。此外，它还可用于研究蛋白质相互作用。本方案详细介绍了获取和解释CD数据的基本步骤，以及分析光谱以估计蛋白质二级结构组成的方法。CD的优点是可以在几小时内对生理缓冲液中含有≤20微克蛋白质的多个样品进行测量。然而，它无法提供可通过X射线晶体学或核磁共振获得的残基特异性信息。
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