Titration_DB: Storage and analysis of NMR-monitored protein pH titration curves
Titration_DB: Storage and analysis of NMR-monitored protein pH titration curves
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DOI：
10.1002/prot.22611
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发表时间：
2010-03-01
影响因子：
2.9
通讯作者：
Nielsen, Jens Erik
中科院分区：
文献类型：
--
作者：
Farrell, Damien;Miranda, Emanuel Sa;Nielsen, Jens Erik
NMR监测的pH滴定实验通常用于测量位点特异性蛋白质pKa值。准确的实验pKa值是必不可少的解剖酶催化，在研究蛋白质的稳定性和配体结合的pH依赖性，在基准pKa预测算法，并最终在理解蛋白质的静电效应。然而，由于pH依赖性静电和结构变化在NMR光谱中表现出来的复杂方式，报告的表观pKa值通常取决于NMR pH滴定曲线的分析方式。因此，重要的是保留原始NMR光谱数据，以便记录和可能的重新解释。我们已经建立了一个数据库的主要NMR pH滴定数据，这是通过一个Web界面访问。在这里，我们报告数据库内容的统计数据，并从全球角度分析数据，为拟合NMR滴定曲线的最佳实践提供指导。Titration_DB可在http://enzyme.ucd.ie/Titration_DB上获得。
NMR-monitored pH titration experiments are routinely used to measure site-specific protein pKa values. Accurate experimental pKa values are essential in dissecting enzyme catalysis, in studying the pH-dependence of protein stability and ligand binding, in benchmarking pKa prediction algorithms, and ultimately in understanding electrostatic effects in proteins. However, due to the complex ways in which pH-dependent electrostatic and structural changes manifest themselves in NMR spectra, reported apparent pKa values are often dependent on the way that NMR pH-titration curves are analyzed. It is therefore important to retain the raw NMR spectroscopic data to allow for documentation and possible reinterpretation. We have constructed a database of primary NMR pH-titration data, which is accessible via a web interface. Here, we report statistics of the database contents and analyze the data with a global perspective to provide guidelines on best practice for fitting NMR titration curves. Titration_DB is available at http://enzyme.ucd.ie/Titration_DB.