DMSO-Quenched H/D-Exchange 2D NMR Spectroscopy and Its Applications in Protein Science

Kunihiro Kuwajima, Maho Yagi-Utsumi, Saeko Yanaka, Koichi Kato

研究成果: ジャーナルへの寄稿総説査読

4 被引用数 (Scopus)

抄録

Hydrogen/deuterium (H/D) exchange combined with two-dimensional (2D) NMR spectroscopy has been widely used for studying the structure, stability, and dynamics of proteins. When we apply the H/D-exchange method to investigate non-native states of proteins such as equilibrium and kinetic folding intermediates, H/D-exchange quenching techniques are indispensable, because the exchange reaction is usually too fast to follow by 2D NMR. In this article, we will describe the dimethylsulfoxide (DMSO)-quenched H/D-exchange method and its applications in protein science. In this method, the H/D-exchange buffer is replaced by an aprotic DMSO solution, which quenches the exchange reaction. We have improved the DMSO-quenched method by using spin desalting columns, which are used for medium exchange from the H/D-exchange buffer to the DMSO solution. This improvement has allowed us to monitor the H/D exchange of proteins at a high concentration of salts or denaturants. We describe methodological details of the improved DMSO-quenched method and present a case study using the improved method on the H/D-exchange behavior of unfolded human ubiquitin in 6 M guanidinium chloride.

本文言語英語
論文番号3748
ジャーナルMolecules
27
12
DOI
出版ステータス出版済み - 6月 1 2022
外部発表はい

!!!All Science Journal Classification (ASJC) codes

  • 分析化学
  • 化学(その他)
  • 分子医療
  • 薬科学
  • 創薬
  • 物理化学および理論化学
  • 有機化学

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