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HNCA+, HNCO+, and HNCACB+ experiments: improved performance by simultaneous detection of orthogonal coherence transfer pathways

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Abstract

Three experiments, BEST–TROSY HNCA+, HNCO+ and HNCACB+ are presented for sequential backbone resonance assignment of 13C, 15N labelled proteins. The novelty of these experiments with respect to conventional pulse sequences is the detection of additional orthogonal coherence transfer pathways that results in enhanced sensitivity for sequential correlations without significantly compromising the intensity of intra-residue correlation peaks. In addition, a 2-step phase cycle separates peaks originating from the orthogonal coherence transfer pathways in 2 sub-spectra, thus providing similar information as obtained from performing a pair of sequential and intra-residue correlation experiments.

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Acknowledgments

We thank Zsofia Solyom for making a sample of NS5A available for this study, and Isabel Ayala for preparation of the ubiquitin sample. This work has been partly supported by grants from the European Union (FP7-I3-BIO-NMR contract No. 261862, FP7-ITN-IDPbyNMR contract No. 264257). S.G. acknowledges support from Bruker Biospin (France). This work used the NMR and isotope labeling platforms of the Grenoble Instruct centre (ISBG; UMS 3518 CNRS-CEA-UJF-EMBL) with support from FRISBI (ANR-10-INSB-05-02) and GRAL (ANR-10-LABX-49-01) within the Grenoble Partnership for Structural Biology (PSB).

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Authors and Affiliations

  1. Institut de Biologie Structurale, Université Grenoble 1, 71 Avenue des Martyrs, 38044, Grenoble Cedex 9, France

    Sergio Gil-Caballero, Adrien Favier & Bernhard Brutscher

  2. Commissariat à l’Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France

    Sergio Gil-Caballero, Adrien Favier & Bernhard Brutscher

  3. Centre National de Recherche Scientifique (CNRS), Grenoble, France

    Sergio Gil-Caballero, Adrien Favier & Bernhard Brutscher

Authors
  1. Sergio Gil-Caballero
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  2. Adrien Favier
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  3. Bernhard Brutscher
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Correspondence to Bernhard Brutscher.

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Gil-Caballero, S., Favier, A. & Brutscher, B. HNCA+, HNCO+, and HNCACB+ experiments: improved performance by simultaneous detection of orthogonal coherence transfer pathways. J Biomol NMR 60, 1–9 (2014). https://doi.org/10.1007/s10858-014-9847-x

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  • DOI: https://doi.org/10.1007/s10858-014-9847-x

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