Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
References
Arntson KE, Pomerantz WCK (2016) Protein-observed fluorine NMR: a bioorthogonal approach for small molecule discovery. J Med Chem 59:5158–5171. https://doi.org/10.1021/acs.jmedchem.5b01447
Bjornson ME, Corson DC, Sykes BD (1985) 13C and 113Cd NMR studies of the chelation of metal ions by the calcium binding protein parvalbumin. J Inorg Biochem 25:141–149. https://doi.org/10.1016/0162-0134(85)80022-2
Boeszoermenyi A, Chhabra S, Dubey A et al (2019) Aromatic 19F- 13C TROSY: a background-free approach to probe biomolecular structure, function, and dynamics. Nat Methods 16:333–340. https://doi.org/10.1038/s41592-019-0334-x
Burt CT (1987) Phosphorus NMR in Biology. CRC Press, Boca Ranton, Fl
Didenko T, Liu JJ, Horst R et al (2013) Fluorine-19 NMR of integral membrane proteins illustrated with studies of GPCRs. Curr Opin Struct Biol 23:740–747. https://doi.org/10.1016/J.SBI.2013.07.011
Fernández C, Wider G (2003) TROSY in NMR studies of the structure and function of large biological macromolecules. Curr Opin Struct Biol 13:570–580. https://doi.org/10.1016/J.SBI.2003.09.009
Griffin RG, Ellett JD, Mehring M et al (1972) Single crystal study of the 19F shielding tensors of a trifluoromethyl group. J Chem Phys 57:2147–2155. https://doi.org/10.1063/1.1678543
Haiech J, Derancourt J, Pechere JF, Demaille JG (1979) A new large-scale purification procedure for muscular parvalbumins. Biochimie 61:583–587. https://doi.org/10.1016/S0300-9084(79)80155-8
Hull WE, Sykes BD (1975) Fluorotyrosine alkaline phosphatase: internal mobility of individual tyrosines and the role of chemical shift anisotropy as a 19F nuclear spin relaxation mechanism in proteins. J Mol Biol 98:121–153. https://doi.org/10.1016/S0022-2836(75)80105-7
Manglik A, Kim TH, Masureel M et al (2015) Structural Insights into the dynamic process of β2-adrenergic receptor signaling. Cell 161:1101–1111. https://doi.org/10.1016/J.CELL.2015.04.043
Matson GB (1977) Methyl NMR relaxation: the effects of spin rotation and chemical shift anisotropy mechanisms. J Chem Phys 67:5152–5161. https://doi.org/10.1063/1.434744
Miclet E, Williams DC, Clore GM et al (2004) Relaxation-optimized NMR spectroscopy of methylene groups in proteins and nucleic acids. J Am Chem Soc 126:10560–10570. https://doi.org/10.1021/ja047904v
Milbrat AG, Arthanari H, Takeuchi K et al (2015) Increased resolution of aromatic cross peaks using alternate 13C labeling and TROSY. J Biomol NMR 62:291–301. https://doi.org/10.1038/nm.2451.A
Naugler D, Prosser RS (2017) Notes on synthesis of perdeutero-5-13C,5,5,5-trifluoroisoleucine VI. BioRxiv. https://doi.org/10.1101/140681
Nelson DJ (1978) Fluorine-19 magnetic resonance of muscle calcium binding parvalbumin: pH dependency of resonance position and spin-lattice relaxation time. Inorg Chim Acta 27:71–74. https://doi.org/10.1016/S0020-1693(00)87233-3
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 94:12366–12371
Prestegard JH, Grant DM (1978) Characterization of anisotropic motion in fatty acid micelles by analysis of transverse relaxation in an AX2 nuclear spin system. J Am Chem Soc 100:4664–4668. https://doi.org/10.1021/ja00483a005
Rashid S, Lee BL, Wajda B, Spyracopoulos L (2019) Side-chain dynamics of the trifluoroacetone cysteine derivative characterized by 19F NMR relaxation and molecular dynamics simulations. J Phys Chem. https://doi.org/10.1021/acs.jpcb.9b01741
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
Seamon KB, Hartshorne DJ, Bothner-By AA (1977) Ca2+ and Mg2+ dependent conformations of troponin C as determined by 1H and 19F nuclear magnetic resonance. Biochemistry 16:4039–4046. https://doi.org/10.1021/bi00637a016
Shimizu H (1964) Theory of the dependence of nuclear magnetic relaxation on the absolute sign of spin-spin coupling constant. J Chem Phys 40:3357–3364. https://doi.org/10.1063/1.1725007
Sykes BD (1969) An application of transient nuclear magnetic resonance methods to the measurement of biological exchange rates. the interaction of trifluoroacetyl-d-phenylalanine with the chymotrypsins. J Am Chem Soc 91:949–955. https://doi.org/10.1021/ja01032a027
Tokunaga Y, Takeuchi K, Shimada I (2017) Forbidden coherence transfer of 19F nuclei to quantitatively measure the dynamics of a CF3-containing ligand in receptor-bound states. Molecules 22:1492–1503. https://doi.org/10.3390/molecules22091492
Tugarinov V, Hwang PM, Ollerenshaw JE, Kay LE (2003) Cross-correlated relaxation enhanced 1H-13C NMR spectroscopy of methyl groups in very high molecular weight proteins and protein complexes. J Am Chem Soc 125:10420–10428. https://doi.org/10.1021/ja030153x
Vold RL, Vold RR (1978) Nuclear magnetic relaxation in coupled spin systems. Prog Nucl Magn Reson Spectrosc 12:79–133. https://doi.org/10.1016/0079-6565(78)80004-1
Werbelow LG, Grant DM (1975) Carbon-13 relaxation in multispin systems of the type AXn. J Chem Phys 63:544–556. https://doi.org/10.1063/1.431085
Acknowledgements
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.
Funding
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.
Author information
Authors and Affiliations
Contributions
BAK, BDS contributed equally to the execution of these experiments and the writing of the paper.
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
10858_2019_266_MOESM1_ESM.pdf
Supporting Information.Protocol for 19F-labelling carp-β parvalbumin, NMR experimental conditions and Supplementary Figures S1-7. (PDF 6131 kb)
Rights and permissions
About this article
Cite this article
Klein, B.A., Sykes, B.D. Feasibility of trifluoromethyl TROSY NMR at high magnetic fields. J Biomol NMR 73, 519–523 (2019). https://doi.org/10.1007/s10858-019-00266-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10858-019-00266-0