Szulik Marta W, Voehler Markus, Stone Michael P
Department of Chemistry and Center for Structural Biology, Vanderbilt University, Nashville, Tennesee; These authors contributed equally to this work.
Curr Protoc Nucleic Acid Chem. 2014 Dec 12;59:7.20.1-18. doi: 10.1002/0471142700.nc0720s59.
Base pairing in nucleic acids plays a crucial role in their structure and function. Differences in the base-pair opening and closing kinetics of individual double-stranded DNA sequences or between chemically modified base pairs provide insight into the recognition of these base pairs by DNA processing enzymes. This unit describes how to quantify the kinetics for localized base pairs by observing changes in the imino proton signals by nuclear magnetic resonance spectroscopy. The determination of all relevant parameters using state-of-the art techniques and NMR instrumentation, including cryoprobes, is discussed.
核酸中的碱基配对在其结构和功能中起着至关重要的作用。单个双链DNA序列或化学修饰碱基对之间碱基对打开和关闭动力学的差异，为深入了解DNA加工酶对这些碱基对的识别提供了线索。本单元介绍了如何通过核磁共振波谱观察亚氨基质子信号的变化来量化局部碱基对的动力学。还讨论了使用包括低温探头在内的先进技术和核磁共振仪器确定所有相关参数的方法。
