A Unique and Simple Approach to Improve Sensitivity in (15)N-NMR Relaxation Measurements for NH?? Groups: Application to a Protein-DNA Complex.
Related Articles A Unique and Simple Approach to Improve Sensitivity in (15)N-NMR Relaxation Measurements for NH?? Groups: Application to a Protein-DNA Complex.
Molecules. 2017 Aug 15;22(8):
Authors: Nguyen D, Lokesh GLR, Volk DE, Iwahara J
Abstract
NMR spectroscopy is a powerful tool for research on protein dynamics. In the past decade, there has been significant progress in the development of NMR methods for studying charged side chains. In particular, NMR methods for lysine side-chain NH?? groups have been proven to be powerful for investigating the dynamics of hydrogen bonds or ion pairs that play important roles in biological processes. However, relatively low sensitivity has been a major practical issue in NMR experiments on NH?? groups. In this paper, we present a unique and simple approach to improve sensitivity in (15)N relaxation measurements for NH?? groups. In this approach, the efficiency of coherence transfers for the desired components are maximized, whereas undesired anti-phase or multi-spin order components are purged through pulse schemes and rapid relaxation. For lysine side-chain NH?? groups of a protein-DNA complex, we compared the data obtained with the previous and new pulse sequences under the same conditions and confirmed that the (15)N relaxation parameters were consistent for these datasets. While retaining accuracy in measuring (15)N relaxation, our new pulse sequences for NH?? groups allowed an 82% increase in detection sensitivity of (15)N longitudinal and transverse relaxation measurements.
PMID: 28809801 [PubMed - in process]
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