Solid-State NMR 19F–1H–15N Correlation Experiments for Resonance Assignment and Distance Measurements of Multifluorinated Proteins
Name
nihms-1965646.pdf
Description
Accepted version
Size
1.96 MB
Format
Adobe PDF
Checksum (MD5)
2ef8599818948dc8ef16c1e668bd23f3
Author(s)
Duan, Pu
Dregni, Aurelio J
Hong, Mei
Date Issued
September 23, 2022
Journal
The Journal of Physical Chemistry A
Publisher
American Chemical Society
Citation
J. Phys. Chem. A 2022, 126, 39, 7021–7032C
Version
Author's final manuscript
Abstract
Several solid-state NMR techniques have been introduced recently to measure nanometer distances involving 19F, whose high gyromagnetic ratio makes it a potent nuclear spin for structural investigation. These solid-state NMR techniques either use 19F correlation with 1H or 13C to obtain qualitative interatomic contacts or use the rotational-echo double-resonance (REDOR) pulse sequence to measure quantitative distances. However, no NMR technique is yet available for disambiguating 1H–19F distances in multiply fluorinated proteins and protein–ligand complexes. Here, we introduce a three-dimensional (3D) 19F–15N–1H correlation experiment that resolves the distances of multiple fluorines to their adjacent amide protons. We show that optimal polarization transfer between 1H and 19F spins is achieved using an out-and-back 1H–19F REDOR sequence. We demonstrate this 3D correlation experiment on the model protein GB1 and apply it to the multidrug-resistance transporter, EmrE, complexed to a tetrafluorinated substrate. This technique should be useful for resolving and assigning distance constraints in multiply fluorinated proteins, leading to significant savings of time and precious samples compared to producing several singly fluorinated samples. Moreover, the method enables structural determination of protein–ligand complexes for ligands that contain multiple fluorines.
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DOI of Published Version
https://doi.org/10.1021/acs.jpca.2c05154
