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Amide additives improve RDC measurements in polyacrylamide.
Journal of Biomolecular NMR ( IF 2.7 ) Pub Date : 2020-02-13 , DOI: 10.1007/s10858-020-00305-1
Talia Fargason 1 , Ting Wang 1 , Naiduwadura Ivon Upekala De Silva 1 , Zihan Zhang 1 , Heather McKelvey 1 , Taylor Knapp 1 , Steve Zaharias 1 , Jun Zhang 1
Affiliation  

Residual dipolar couplings (RDCs) provide valuable NMR parameters that can be used for structural calculation and verification. Measuring RDCs requires aligning macromolecules using one of various types of alignment media. Of different alignment media options, stretched or compressed polyacrylamide gels are advantageous due to their chemical stability. However, polyacrylamide interacts with proteins and significantly broadens NMR resonances. In this study, we found that the amide-containing compounds asparagine, glutamine and propionamide improve spectral quality of proteins in polyacrylamide gel without significantly reducing the magnitude of RDC values. Moreover, we showed that propionamide is an attractive additive that increases protein solubility without interfering with protein stability, ligand binding or NMR pulse width, suggesting its potential applications for our NMR methods.

中文翻译:

酰胺添加剂可改善聚丙烯酰胺中的RDC测量。

残留偶极耦合(RDC)提供了可用于结构计算和验证的有价值的NMR参数。测量RDC需要使用多种类型的校准介质之一来校准大分子。在不同的取向介质选择中,拉伸或压缩的聚丙烯酰胺凝胶由于其化学稳定性而具有优势。但是,聚丙烯酰胺与蛋白质相互作用并显着拓宽NMR共振。在这项研究中,我们发现含酰胺的化合物天冬酰胺,谷氨酰胺和丙酰胺可改善聚丙烯酰胺凝胶中蛋白质的光谱质量,而不会显着降低RDC值的大小。此外,我们发现丙酰胺是一种有吸引力的添加剂,可以增加蛋白质溶解度而不会干扰蛋白质稳定性,配体结合或NMR脉冲宽度,
更新日期:2020-04-21
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