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

Journal of Biomolecular NMR
Koh TakeuchiIchio Shimada

Abstract

Direct detection of the TROSY component of proton-attached (15)N nuclei ((15)N-detected TROSY) yields high quality spectra with high field magnets, by taking advantage of the slow (15)N transverse relaxation. The slow transverse relaxation and narrow line width of the (15)N-detected TROSY resonances are expected to compensate for the inherently low (15)N sensitivity. However, the sensitivity of (15)N-detected TROSY in a previous report was one-order of magnitude lower than in the conventional (1)H-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 (1)H-detected experiments. Here, we show that the sensitivity gap between (15)N and (1)H 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 (15)N-detected TROSY. Together with the previously reported benefits of the (15)N-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.

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Citations

Mar 2, 2016·Protein Science : a Publication of the Protein Society·Kirill Oxenoid, James J Chou
May 16, 2017·Archives of Biochemistry and Biophysics·Eric B GibbsScott A Showalter
Sep 8, 2017·Journal of Biomolecular NMR·Robbin SchniedersBoris Fürtig
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Feb 13, 2018·Proceedings of the National Academy of Sciences of the United States of America·Sandeep ChhabraHaribabu Arthanari
Nov 21, 2016·Journal of Biomolecular NMR·Koh TakeuchiGerhard Wagner
Mar 12, 2020·International Journal of Molecular Sciences·Yuji TokunagaKoh Takeuchi
Dec 15, 2020·Protein Science : a Publication of the Protein Society·Kathleen F O'RourkeDavid D Boehr
Dec 31, 2020·International Journal of Molecular Sciences·Yuji Tokunaga, Koh Takeuchi
Jun 4, 2019·Journal of Inorganic Biochemistry·Samah Al-HarthiŁukasz Jaremko
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Dec 31, 2019·Journal of the American Chemical Society·Kyle W EastGeorge P Lisi
Oct 12, 2021·Journal of the American Chemical Society·Gogulan KarunanithyD Flemming Hansen

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