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BEST and SOFAST experiments for resonance assignment of histidine and tyrosine side chains in 13C/15N labeled proteins

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Abstract

Aromatic amino-acid side chains are essential components for the structure and function of proteins. We present herein a set of NMR experiments for time-efficient resonance assignment of histidine and tyrosine side chains in uniformly 13C/15N-labeled proteins. The use of band-selective 13C pulses allows to deal with linear chains of coupled spins, thus avoiding signal loss that occurs in branched spin systems during coherence transfer. Furthermore, our pulse schemes make use of longitudinal 1H relaxation enhancement, Ernst-angle excitation, and simultaneous detection of 1H and 13C steady-state polarization to achieve significant signal enhancements.

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Acknowledgements

We thank Dominique Bourgeois and Virgile Adam for making the rsfolder plasmid available, and Isabel Ayala and Karine Giandoreggio for protein sample preparation. We acknowledge support from the CNRS (Défis Instrumentation 2018). This work used the NMR and isotope labeling platforms of the Grenoble Instruct center (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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  1. Univ. Grenoble Alpes, CEA, CNRS, IBS, 38000, Grenoble, France

    Nina Eleni Christou & Bernhard Brutscher

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  1. Nina Eleni Christou
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  2. Bernhard Brutscher
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Correspondence to Bernhard Brutscher.

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Christou, N.E., Brutscher, B. BEST and SOFAST experiments for resonance assignment of histidine and tyrosine side chains in 13C/15N labeled proteins. J Biomol NMR 72, 115–124 (2018). https://doi.org/10.1007/s10858-018-0216-z

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  • DOI: https://doi.org/10.1007/s10858-018-0216-z

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