Nitrogen-detected TROSY yields comparable sensitivity to proton-detected TROSY for non-deuterated, large proteins under physiological salt conditions

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• 作者：Koh Takeuchi ; Haribabu Arthanari ; Misaki Imai…
• 关键词：Nitrogen detection ; TROSY ; High field magnet ; Protein NMR ; Salt concentration ; Sensitivity
• 刊名：Journal of Biomolecular NMR
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：64
• 期：2
• 页码：143-151
• 全文大小：1,473 KB
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• 作者单位：Koh Takeuchi (1) (2)
  Haribabu Arthanari (3)
  Misaki Imai (4)
  Gerhard Wagner (3)
  Ichio Shimada (1) (5)
  
  1. Molecular Profiling Research Center for Drug Discovery, National Institute for Advanced Industrial Science and Technology, Tokyo, 135-0063, Japan
  2. PRESTO, JST, Tokyo, 135-0063, Japan
  3. Department of Biochemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA, 02115, USA
  4. Research and Development Department, Japan Biological Informatics Consortium, Tokyo, 135-0063, Japan
  5. Graduate Schools of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan
  
• 刊物类别：Physics and Astronomy
• 刊物主题：Physics
  Biophysics and Biomedical Physics
  Polymer Sciences
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-5001

文摘

Direct detection of the TROSY component of proton-attached 15N nuclei (15N-detected TROSY) yields high quality spectra with high field magnets, by taking advantage of the slow 15N transverse relaxation. The slow transverse relaxation and narrow line width of the 15N-detected TROSY resonances are expected to compensate for the inherently low 15N sensitivity. However, the sensitivity of 15N-detected TROSY in a previous report was one-order of magnitude lower than in the conventional 1H-detected version. This could be due to the fact that the previous experiments were performed at low salt (0–50 mM), which is advantageous for 1H-detected experiments. Here, we show that the sensitivity gap between 15N and 1H becomes marginal for a non-deuterated, large protein (τ _c = 35 ns) at a physiological salt concentration (200 mM). This effect is due to the high salt tolerance of the 15N-detected TROSY. Together with the previously reported benefits of the 15N-detected TROSY, our results provide further support for the significance of this experiment for structural studies of macromolecules when using high field magnets near and above 1 GHz. Keywords Nitrogen detection TROSY High field magnet Protein NMR Salt concentration Sensitivity