一种用于在蛋白质 - 核酸复合物中对形成分子间离子对的蛋白质侧链NH₃⁺基团的核磁共振信号进行位点特异性识别的化学方法。
A chemical approach for site-specific identification of NMR signals from protein side-chain NH₃⁺ groups forming intermolecular ion pairs in protein-nucleic acid complexes.
作者信息
Anderson Kurtis M, Nguyen Dan, Esadze Alexandre, Zandrashvili Levani, Gorenstein David G, Iwahara Junji
机构信息
Department of NanoMedicine and Biomedical Engineering and Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, TX, 77225, USA.
出版信息
J Biomol NMR. 2015 May;62(1):1-5. doi: 10.1007/s10858-015-9909-8. Epub 2015 Feb 19.
Abstract
Protein-nucleic acid interactions involve intermolecular ion pairs of protein side-chain and DNA or RNA phosphate groups. Using three protein-DNA complexes, we demonstrate that site-specific oxygen-to-sulfur substitution in phosphate groups allows for identification of NMR signals from the protein side-chain NH3 (+) groups forming the intermolecular ion pairs. A characteristic change in their (1)H and (15)N resonances upon this modification (i.e., substitution of phosphate to phosphorodithioate) can represent a signature of an intermolecular ion pair. Hydrogen-bond scalar coupling between protein side-chain (15)N and DNA phosphorodithiaote (31)P nuclei provides direct confirmation of the intermolecular ion pair. The same approach is likely applicable to protein-RNA complexes as well.
摘要
蛋白质 - 核酸相互作用涉及蛋白质侧链与DNA或RNA磷酸基团之间的分子间离子对。通过使用三种蛋白质 - DNA复合物，我们证明磷酸基团中位点特异性的氧到硫取代能够识别来自形成分子间离子对的蛋白质侧链NH3(+)基团的NMR信号。这种修饰（即磷酸酯取代为二硫代磷酸酯）后，其(1)H和(15)N共振的特征性变化可代表分子间离子对的特征。蛋白质侧链(15)N与DNA二硫代磷酸酯(31)P核之间的氢键标量耦合直接证实了分子间离子对的存在。同样的方法可能也适用于蛋白质 - RNA复合物。
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