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A comparison of chemical shift sensitivity of trifluoromethyl tags: optimizing resolution in 19F NMR studies of proteins

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Abstract

The elucidation of distinct protein conformers or states by fluorine (19F) NMR requires fluorinated moieties whose chemical shifts are most sensitive to subtle changes in the local dielectric and magnetic shielding environment. In this study we evaluate the effective chemical shift dispersion of a number of thiol-reactive trifluoromethyl probes [i.e. 2-bromo-N-(4-(trifluoromethyl)phenyl)acetamide (BTFMA), N-(4-bromo-3-(trifluoromethyl)phenyl)acetamide (3-BTFMA), 3-bromo-1,1,1-trifluoropropan-2-ol (BTFP), 1-bromo-3,3,4,4,4-pentafluorobutan-2-one (BPFB), 3-bromo-1,1,1-trifluoropropan-2-one (BTFA), and 2,2,2-trifluoroethyl-1-thiol (TFET)] under conditions of varying polarity. In considering the sensitivity of the 19F NMR chemical shift to the local environment, a series of methanol/water mixtures were prepared, ranging from relatively non-polar (MeOH:H2O = 4) to polar (MeOH:H2O = 0.25). 19F NMR spectra of the tripeptide, glutathione ((2S)-2-amino-4-{[(1R)-1-[(carboxymethyl)carbamoyl]-2-sulfanylethyl]carbamoyl}butanoic acid), conjugated to each of the above trifluoromethyl probes, revealed that the BTFMA tag exhibited a significantly greater range of chemical shift as a function of solvent polarity than did either BTFA or TFET. DFT calculations using the B3LYP hybrid functional and the 6-31G(d,p) basis set, confirmed the observed trend in chemical shift dispersion with solvent polarity.

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Acknowledgments

We would like to thank the Natural Sciences and Engineering Research Council of Canada for support of this research. Supported by NSERC research discovery award (Grant No. 261980).

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    Authors and Affiliations

    1. Department of Chemistry, University of Toronto, UTM, 3359 Mississauga Road North, Mississauga, ON, L5L 1C6, Canada

      Libin Ye, Sacha Thierry Larda, Yi Feng Frank Li & R. Scott Prosser

    2. Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, 94305, USA

      Aashish Manglik

    3. Department of Biochemistry, University of Toronto, 1 King’s College Circle, Toronto, ON, M5S 1A8, Canada

      R. Scott Prosser

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    1. Libin Ye
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    2. Sacha Thierry Larda
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    Correspondence to R. Scott Prosser.

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    Libin Ye and Sacha Thierry Larda have contributed equally to this work.

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    Ye, L., Larda, S.T., Frank Li, Y.F. et al. A comparison of chemical shift sensitivity of trifluoromethyl tags: optimizing resolution in 19F NMR studies of proteins. J Biomol NMR 62, 97–103 (2015). https://doi.org/10.1007/s10858-015-9922-y

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    • DOI: https://doi.org/10.1007/s10858-015-9922-y

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