Selectively dispersed isotope labeling for protein structure determination by magic angle spinning NMR
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Author(s)
Eddy, Matthew Thomas
Belenky, Marina
Sivertsen, Astrid C.
Herzfeld, Judith
Griffin, Robert Guy
Date Issued
August 2013
Journal
Journal of Biomolecular NMR
Publisher
Springer-Verlag
Citation
Eddy, Matthew T., Marina Belenky, Astrid C. Sivertsen, Robert G. Griffin, and Judith Herzfeld. “Selectively Dispersed Isotope Labeling for Protein Structure Determination by Magic Angle Spinning NMR.” Journal of Biomolecular NMR 57, no. 2 (August 30, 2013): 129–139.
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Author's final manuscript
Abstract
The power of nuclear magnetic resonance spectroscopy derives from its site-specific access to chemical, structural and dynamic information. However, the corresponding multiplicity of interactions can be difficult to tease apart. Complimentary approaches involve spectral editing on the one hand and selective isotope substitution on the other. Here we present a new “redox” approach to the latter: acetate is chosen as the sole carbon source for the extreme oxidation numbers of its two carbons. Consistent with conventional anabolic pathways for the amino acids, [1-[superscript 13]C] acetate does not label α carbons, labels other aliphatic carbons and the aromatic carbons very selectively, and labels the carboxyl carbons heavily. The benefits of this labeling scheme are exemplified by magic angle spinning spectra of microcrystalline immunoglobulin binding protein G (GB1): the elimination of most J-couplings and one- and two-bond dipolar couplings provides narrow signals and long-range, intra- and inter-residue, recoupling essential for distance constraints. Inverse redox labeling, from [2-[superscript 13]C] acetate, is also expected to be useful: although it retains one-bond couplings in the sidechains, the removal of CA–CO coupling in the backbone should improve the resolution of NCACX spectra.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
Francis Bitter Magnet Laboratory (Massachusetts Institute of Technology)
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DOI of Published Version
https://doi.org/10.1007/s10858-013-9773-3
