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High-resolution aliphatic side-chain assignments in 3D HCcoNH experiments with joint H–C evolution and non-uniform sampling

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Abstract

We describe an efficient NMR triple resonance approach that correlates, at high resolution, protein side-chain and backbone resonances. It relies on the combination of two strategies: joint evolution of aliphatic side-chain proton/carbon coherences using a backbone N–H based HCcoNH reduced dimensionality (RD) experiment and non-uniform sampling (NUS) in two indirect dimensions. A typical data set containing such correlation information can be acquired in 2 days, at very high resolution unfeasible for conventional 4D HCcoNH-TOCSY experiments. The resonances of the aliphatic side-chain protons are unambiguously assigned to their attached carbons through the analysis of the ‘sum’ and ‘difference’ spectra. This approach circumvents the tedious process of manual resonance assignments using HCcH-TOCSY data, while providing additional resolving power of backbone N–H signals. A simple peak-list based algorithm has been implemented in the IBIS software for rapid automated backbone and side-chain assignments.

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Authors and Affiliations

  1. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 200 Longwood Avenue, 02115, Boston, MA, USA

    Zhen-Yu J. Sun, Sven G. Hyberts, David Rovnyak, Sunghyouk Park & Gerhard Wagner

  2. Department of Chemistry, Bucknell University, 17837, Lewisburg, PA, USA

    David Rovnyak

  3. Rowland Institute at Harvard, 02142, Cambridge, MA, USA

    Alan S. Stern

  4. Department of Molecular, Microbial, and Structural Biology, University of Connecticut Health Center, 06030, Farmington, CT, USA

    Jeffrey C. Hoch

Authors
  1. Zhen-Yu J. Sun
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  2. Sven G. Hyberts
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  3. David Rovnyak
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  4. Sunghyouk Park
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  5. Alan S. Stern
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  6. Jeffrey C. Hoch
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  7. Gerhard Wagner
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Correspondence to Gerhard Wagner.

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Sun, ZY.J., Hyberts, S.G., Rovnyak, D. et al. High-resolution aliphatic side-chain assignments in 3D HCcoNH experiments with joint H–C evolution and non-uniform sampling. J Biomol NMR 32, 55–60 (2005). https://doi.org/10.1007/s10858-005-3339-y

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  • DOI: https://doi.org/10.1007/s10858-005-3339-y

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