Koscielniak, JanuszLi, JessSail, DeepakSwenson, RolfAnklin, ClemensRozovsky, SharonByrd, R. Andrew2024-02-092024-02-092023-11-15Koscielniak, Janusz, Jess Li, Deepak Sail, Rolf Swenson, Clemens Anklin, Sharon Rozovsky, and R. Andrew Byrd. “Exploring Sulfur Sites in Proteins via Triple-Resonance 1H-Detected 77Se NMR.” Journal of the American Chemical Society 145, no. 45 (November 15, 2023): 24648–56. https://doi.org/10.1021/jacs.3c07225.1520-5126https://udspace.udel.edu/handle/19716/33966This article was originally published in Journal of the American Chemical Society. The version of record is available at: https://doi.org/10.1021/jacs.3c07225. Copyright © 2023 The Authors. Published by American Chemical Society.NMR spectroscopy has been applied to virtually all sites within proteins and biomolecules; however, the observation of sulfur sites remains very challenging. Recent studies have examined 77Se as a replacement for sulfur and applied 77Se NMR in both the solution and solid states. As a spin-1/2 nuclide, 77Se is attractive as a probe of sulfur sites, and it has a very large chemical shift range (due to a large chemical shift anisotropy), which makes it potentially very sensitive to structural and/or binding interactions as well as dynamics. Despite being a spin-1/2 nuclide, there have been rather limited studies of 77Se, and the ability to use 1H-indirect detection has been sparse. Some examples exist, but in the absence of a directly bonded, nonexchangeable 1H, these have been largely limited to smaller molecules. We develop and illustrate approaches using double-labeling of 13C and 77Se in proteins that enable more sensitive triple-resonance schemes via multistep coherence transfers and 1H-detection. These methods require specialized hardware and decoupling schemes, which we developed and will be discussed.en-USAttribution-NonCommercial-NoDerivatives 4.0 InternationalArticle