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

被引:12
作者
Anderson, Kurtis M. [1 ]
Nguyen, Dan [2 ]
Esadze, Alexandre [2 ]
Zandrashvili, Levani [2 ]
Gorenstein, David G. [1 ]
Iwahara, Junji [2 ]
机构
[1] Univ Texas Hlth Sci Ctr Houston, Inst Mol Med, Dept NanoMed & Biomed Engn, Houston, TX 77225 USA
[2] Univ Texas Med Branch, Sealy Ctr Struct Biol & Mol Biophys, Dept Biochem & Mol Biol, Galveston, TX 77555 USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
Protein-nucleic acid interactions; Protein side chains; NH3+ groups; Ion pairs; Hydrogen bonds; LYSINE; DNA; PHOSPHOROTHIOATE; SEPARATION; DYNAMICS; N-15; SPECTROSCOPY;
D O I
10.1007/s10858-015-9909-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 H-1 and N-15 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 N-15 and DNA phosphorodithiaote P-31 nuclei provides direct confirmation of the intermolecular ion pair. The same approach is likely applicable to protein-RNA complexes as well.
引用
收藏
页码:1 / 5
页数:5
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