文章基本信息

标题：Detecting O2 binding sites in protein cavities
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作者：Ryo Kitahara ; Yuichi Yoshimura ; Mengjun Xue 等
期刊名称：Scientific Reports
电子版ISSN：2045-2322
出版年度：2016
卷号：6
期号：1
DOI：10.1038/srep20534
语种：English
出版社：Springer Nature
摘要：Internal cavities are important elements in protein structure, dynamics, stability and function. Here we use NMR spectroscopy to investigate the binding of molecular oxygen (O₂) to cavities in a well-studied model for ligand binding, the L99A mutant of T4 lysozyme. On increasing the O₂ concentration to 8.9 mM, changes in ¹H, ¹⁵N, and ¹³C chemical shifts and signal broadening were observed specifically for backbone amide and side chain methyl groups located around the two hydrophobic cavities of the protein. O₂-induced longitudinal relaxation enhancements for amide and methyl protons could be adequately accounted for by paramagnetic dipolar relaxation. These data provide the first experimental demonstration that O₂ binds specifically to the hydrophobic, and not the hydrophilic cavities, in a protein. Molecular dynamics simulations visualized the rotational and translational motions of O₂ in the cavities, as well as the binding and egress of O₂, suggesting that the channel consisting of helices D, E, G, H, and J could be the potential gateway for ligand binding to the protein. Due to strong paramagnetic relaxation effects, O₂ gas-pressure NMR measurements can detect hydrophobic cavities when populated to as little as 1%, and thereby provide a general and highly sensitive method for detecting oxygen binding in proteins.