Feasibility of trifluoromethyl TROSY NMR at high magnetic fields
- 295 Downloads
Insights into the structure and dynamics of large biological systems has been greatly improved by two concurrent NMR approaches: the application of transverse relaxation-optimized spectroscopy (TROSY) techniques in multi-dimensional NMR, especially the methyl-TROSY, and the resurgence of 19F NMR using trifluoromethyl (CF3) probes. Herein we investigate the feasibility of combining these approaches into a trifluoromethyl-TROSY experiment. Using a CF3-labelled parvalbumin, we have evaluated the natural abundance 13C-19F correlation spectra and find no indication of a CF3 TROSY at high magnetic fields.
KeywordsMethyl-TROSY Trifluoromethyl 19F NMR Chemical shift anisotropy High magnetic fields
We would like to thank Dr. Jack Moore of the Alberta Proteomics and Mass Spectrometry Facility for assistance with mass spectrometry of proteins and Dr. Andrew Atkinson at King’s College London for 400 MHz NMR spectra.
BAK, BDS contributed equally to the execution of these experiments and the writing of the paper.
This research has been supported by grants from The Heart and Stroke Foundation of Canada (G-14-0005884, BDS) and Faculty of Medicine Transitional Program.
- Burt CT (1987) Phosphorus NMR in Biology. CRC Press, Boca Ranton, FlGoogle Scholar
- Pervushin K, Riek R, Wider G, Wu¨thrich KW (1997) Attenuated T2 relaxation by mutual cancellation of dipole-dipole coupling and chemical shift anisotropy indicates an avenue to NMR structures of very large biological macromolecules in solution. Proc Natl Acad Sci 94:12366–12371ADSCrossRefGoogle Scholar
- Rosenzweig R, Kay LE (2014) bringing dynamic molecular machines into focus by methyl-TROSY NMR. Annu Rev Biochem 83:291–315. https://doi.org/10.1146/annurev-biochem-060713-035829 CrossRefGoogle Scholar