Journal
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volume 143, Issue 46, Pages 19587-19598Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jacs.1c10104
Keywords
-
Categories
Funding
- Australian Research Council [FL170100019, DP200100348, DP21010088, CE200100012]
- Australian Research Council [FL170100019, CE200100012, DP200100348] Funding Source: Australian Research Council
Ask authors/readers for more resources
In this study, it was demonstrated that fluorine atoms in noncanonical amino acids positioned in the hydrophobic core or on the surface of a protein can be linked by scalar through-space F-19-F-19 couplings, even if the F-19 spins are separated by more than the van der Waals distance. The use of fluorinated amino acids and F-19-F-19 interactions offer a straightforward way to probe the spatial proximity of selected sites in proteins, without requiring H-1 NMR resonance assignments.
Fluorine atoms are known to display scalar F-19-F-19 couplings in nuclear magnetic resonance (NMR) spectra when they are sufficiently close in space for nonbonding orbitals to overlap. We show that fluorinated noncanonical amino acids positioned in the hydrophobic core or on the surface of a protein can be linked by scalar through-space F-19-F-19 ((TS)J(FF)) couplings even if the F-19 spins are in the time average separated by more than the van der Waals distance. Using two different aromatic amino acids featuring CF3 groups, O-trifluoromethyl-tyrosine and 4-trifluoromethyl-phenylalanine, we show that F-19-F-19 TOCSY experiments are sufficiently sensitive to detect (TS)J(FF) couplings between 2.5 and 5 Hz in the 19 kDa protein PpiB measured on a two-channel 400 MHz NMR spectrometer with a regular room temperature probe. A quantitative J evolution experiment enables the measurement of (TS)J(FF) coupling constants that are up to five times smaller than the F-19 NMR line width. In addition, a new aminoacyl-tRNA synthetase was identified for genetic encoding of N-6-(trifluoroacetyl)-L-lysine (TFA-Lys) and F-19-F-19 TOCSY peaks were observed between two TFA-Lys residues incorporated into the proteins AncCDT-1 and mRFP despite high solvent exposure and flexibility of the TFA-Lys side chains. With the ready availability of systems for site-specific incorporation of fluorinated amino acids into proteins by genetic encoding, F-19-F-19 interactions offer a straightforward way to probe the spatial proximity of selected sites without any assignments of H-1 NMR resonances.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available