Rapid measurement of long-range distances in proteins by multidimensional 13C-19F REDOR NMR under fast magic-angle spinning

The ability to simultaneously measure many long-range distances is critical to efficient and accurate determination of protein structures by solid-state NMR (SSNMR). So far, the most common distance constraints for proteins are [subscript 13]C-[subscript 15]N distances, which are usually measured us...

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Bibliographic Details
Main Authors: Shcherbakov, Alexander Aleksandrovich (Author), Hong, Mei (Author)
Other Authors: Massachusetts Institute of Technology. Department of Chemistry (Contributor)
Format: Article
Language:English
Published: Springer Nature, 2020-06-23T18:01:48Z.
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Online Access:Get fulltext
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042 |a dc 
100 1 0 |a Shcherbakov, Alexander Aleksandrovich  |e author 
100 1 0 |a Massachusetts Institute of Technology. Department of Chemistry  |e contributor 
700 1 0 |a Hong, Mei  |e author 
245 0 0 |a Rapid measurement of long-range distances in proteins by multidimensional 13C-19F REDOR NMR under fast magic-angle spinning 
246 3 3 |a Rapid measurement of long-range distances in proteins by multidimensional [subscript 13]C-[subscript 19]F REDOR NMR under fast magic-angle spinning 
260 |b Springer Nature,   |c 2020-06-23T18:01:48Z. 
856 |z Get fulltext  |u https://hdl.handle.net/1721.1/125939 
520 |a The ability to simultaneously measure many long-range distances is critical to efficient and accurate determination of protein structures by solid-state NMR (SSNMR). So far, the most common distance constraints for proteins are [subscript 13]C-[subscript 15]N distances, which are usually measured using the rotational-echo double-resonance (REDOR) technique. However, these measurements are restricted to distances of up to ~ 5 Å due to the low gyromagnetic ratios of [subscript 15]N and [subscript 13]C. Here we present a robust 2D [subscript 13]C-[subscript 19]F REDOR experiment to measure multiple distances to ~ 10 Å. The technique targets proteins that contain a small number of recombinantly or synthetically incorporated fluorines. The [subscript 13]C-19F REDOR sequence is combined with 2D [subscript 13]C-[subscript 13]C correlation to resolve multiple distances in highly [subscript 13]C-labeled proteins. We show that, at the high magnetic fields which are important for obtaining well resolved [subscript 13]C spectra, the deleterious effect of the large [subscript 19]F chemical shift anisotropy for REDOR is ameliorated by fast magic-angle spinning and is further taken into account in numerical simulations. We demonstrate this 2D [subscript 13]C-[subscript 13]C resolved [subscript 13]C-[subscript 19]F REDOR technique on [subscript 13]C, [subscript 15]N-labeled GB1. A 5[superscript -19]F-Trp tagged GB1 sample shows the extraction of distances to a single fluorine atom, while a [subscript 3-19]F-Tyr labeled GB1 sample allows us to evaluate the effects of multi-spin coupling and statistical [subscript 19]F labeling on distance measurement. Finally, we apply this 2D REDOR experiment to membrane-bound influenza BM2 transmembrane peptide, and show that the distance between the proton-selective histidine residue and the gating tryptophan residue differs from the distances in the solution NMR structure of detergent-bound BM2. This 2D [subscript 13]C-[subscript 19]F REDOR technique should facilitate SSNMR-based protein structure determination by increasing the measurable distances to the ~ 10 Å range. 
520 |a National Institutes of Health (grant nos. GM066976 and GM088204) 
546 |a en 
655 7 |a Article 
773 |t 10.1007/S10858-018-0187-0 
773 |t Journal of Biomolecular NMR