Integrating <sup>19</sup>F Distance Restraints for Accurate Protein Structure Determination by Magic Angle Spinning NMR Spectroscopy
Traditional protein structure determination by magic angle spinning (MAS) solid-state NMR spectroscopy primarily relies on interatomic distances up to 8 Å, extracted from <sup>13</sup>C-, <sup>15</sup>N-, and <sup>1</sup>H-based dipolar-based correlation experiments. Here, we show that <sup>19</sup>F fast (60 kHz) MAS NMR spectroscopy can supply additional, longer distances. Using 4F-Trp,U-<sup>13</sup>C,<sup>15</sup>N crystalline <i>Oscillatoria agardhii</i> agglutinin (OAA), we demonstrate that judiciously designed 2D and 3D <sup>19</sup>F-based dipolar correlation experiments such as (H)CF, (H)CHF, and FF can yield interatomic distances in the 8-16 Å range. Incorporation of fluorine-based restraints into structure calculation improved the precision of Trp side chain conformations as well as regions in the protein around the fluorine containing residues, with notable improvements observed for residues in proximity to the Trp pairs (W10/W17 and W77/W84) in the carbohydrate-binding loops, which lacked sufficient long-range <sup>13</sup>C-<sup>13</sup>C distance restraints. Our work highlights the use of fluorine and <sup>19</sup>F fast MAS NMR spectroscopy as a powerful structural biology tool.
Authors
- Santiago Antolínez (ORCID: https://orcid.org/0009-0006-9415-9873)
- In‐Ja L. Byeon (ORCID: https://orcid.org/0000-0002-8444-0606)
- Angela M. Gronenborn (ORCID: https://orcid.org/0000-0001-9072-3525)
- Ryan W. Russell (ORCID: https://orcid.org/0000-0002-3599-133X)
- Brent Runge (ORCID: https://orcid.org/0000-0003-3759-5143)
- Jochem Struppe (ORCID: https://orcid.org/0000-0001-9001-5991)
- Jodi A. Hadden‐Perilla (ORCID: https://orcid.org/0000-0003-4685-8291)
- Tatyana Polenova (ORCID: https://orcid.org/0000-0002-0346-1131)
- Manman Lu (ORCID: https://orcid.org/0000-0002-4156-4975)
- Charles D. Schwieters (ORCID: https://orcid.org/0000-0002-4216-4658)
- Roman Zadorozhnyi (ORCID: https://orcid.org/0000-0003-2767-2002)
- Caitlin M. Quinn (ORCID: https://orcid.org/0000-0001-6172-7324)
Institutions
- National Institutes of Health (US)
- University of Pittsburgh (US)
- Bruker (United States) (US)
- National Institute of Diabetes and Digestive and Kidney Diseases (US)
- University of Delaware (US)
Publication Details
- Journal
- Journal of the American Chemical Society
- Published
- 2024-10-23
- DOI
- https://doi.org/10.1021/jacs.4c11373
- Citations
- 5
- Primary Topic
- Advanced NMR Techniques and Applications
- Type
- article
- Field-Weighted Citation Impact
- 1.20
Funders
- National Science Foundation
- National Institutes of Health
- National Institute of Allergy and Infectious Diseases
- National Institute of Diabetes and Digestive and Kidney Diseases
- Division of Chemistry