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Anomalous Properties of Lys Residues Buried in the Hydrophobic Interior of a Protein Revealed with 15N-Detect NMR Spectroscopy
The Journal of Physical Chemistry Letters ( IF 5.7 ) Pub Date : 2018-01-09 00:00:00 , DOI: 10.1021/acs.jpclett.7b02668
Christos M. Kougentakis , Emily M. Grasso , Aaron C. Robinson , José A. Caro , Jamie L. Schlessman 1 , Ananya Majumdar , Bertrand García-Moreno E.
Affiliation  

Ionizable residues buried in hydrophobic environments in proteins are essential for many fundamental biochemical processes. These residues titrate with anomalous pKa values that are challenging to reproduce with structure-based calculations owing to the conformational reorganization coupled to their ionization. Detailed characterization of this conformational reorganization is of interest; unfortunately, the properties of buried Lys residues are difficult to study experimentally. Here we demonstrate the utility of 15N NMR spectroscopy to gain insight into the protonation state, state of hydration and conformational dynamics of the Nζ amino group of buried Lys residues. The experiments were applied to five variants of staphylococcal nuclease, with internal Lys residues that titrate with pKa values ranging from 6.2 to 8.1. Direct detection of buried Lys residues with these NMR spectroscopy methods will enable correlation between thermodynamic and structural data as well as unprecedented examination of how conformational transitions coupled to their ionization affect their pKa values.

中文翻译:

15 N检测NMR光谱揭示的蛋白质疏水内部掩埋的Lys残基的异常性质

埋在蛋白质疏水性环境中的可电离残基对于许多基本的生化过程至关重要。这些残基具有异常的p K a滴定度,由于构象重组和其电离作用,基于结构的计算很难重现。这种构象重组的详细表征令人感兴趣。不幸的是,难以通过实验研究掩埋的Lys残基的性质。这里我们展示了15的效用N NMR光谱可深入了解被掩埋的Lys残基的Nζ氨基的质子化状态,水合状态和构象动力学。将实验应用于五种葡萄球菌核酸酶变体,其内部Lys残基滴定的p K a值为6.2至8.1。使用这些NMR光谱法直接检测掩埋的Lys残基将使热力学数据与结构数据之间具有相关性,并且前所未有地研究了构象转变与其电离耦合如何影响其p K a值。
更新日期:2018-01-09
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