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A NMR experiment for simultaneous correlations of valine and leucine/isoleucine methyls with carbonyl chemical shifts in proteins

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Abstract

A methyl-detected ‘out-and-back’ NMR experiment for obtaining simultaneous correlations of methyl resonances of valine and isoleucine/leucine residues with backbone carbonyl chemical shifts, SIM-HMCM(CGCBCA)CO, is described. The developed pulse-scheme serves the purpose of convenience in recording a single data set for all Ileδ1, Leuδ and Valγ (ILV) methyl positions instead of acquiring two separate spectra selective for valine or leucine/isoleucine residues. The SIM-HMCM(CGCBCA)CO experiment can be used for ILV methyl assignments in moderately sized protein systems (up to ~100 kDa) where the backbone chemical shifts of 13Cα, 13Cβ and 13CO are known from prior NMR studies and where some losses in sensitivity can be tolerated for the sake of an overall reduction in NMR acquisition time.

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Acknowledgments

This work was supported by funds from the Intramural Program of the NIH, NIDDK, and the Intramural AIDS Targeted Antiviral Program of the Office of the Director of the NIH (to G.M.C.).

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Authors and Affiliations

  1. Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892-0520, USA

    Vitali Tugarinov, Vincenzo Venditti & G. Marius Clore

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  1. Vitali Tugarinov
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  2. Vincenzo Venditti
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  3. G. Marius Clore
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Correspondence to Vitali Tugarinov or G. Marius Clore.

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10858_2013_9803_MOESM1_ESM.pdf

Supporting Information Available: Listing of the Bruker pulse code for the SIM-HMCM(CGCBCA)CO experiment. NMR acquisition parameters and nmrPipe processing scripts are available from the authors upon request (PDF 83 kb)

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Tugarinov, V., Venditti, V. & Marius Clore, G. A NMR experiment for simultaneous correlations of valine and leucine/isoleucine methyls with carbonyl chemical shifts in proteins. J Biomol NMR 58, 1–8 (2014). https://doi.org/10.1007/s10858-013-9803-1

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  • DOI: https://doi.org/10.1007/s10858-013-9803-1

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