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Pinpoint analysis of a protein in slow exchange using F1F2-selective ZZ-exchange spectroscopy: assignment and kinetic analysis

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Abstract

ZZ-exchange spectroscopy is widely used to study slow exchange processes in biomolecules, especially determination of exchange rates and assignment of minor peaks. However, if the exchange cross peaks overlap or the populations are skewed, kinetic analysis is hindered. In order to analyze slow exchange protein dynamics under such conditions, here we have developed a new method by combining ZZ-exchange and F1F2-selective NMR spectroscopy. We demonstrate the utility of this method by examining the monomer–dimer transition of the ubiquitin-associated domain of p62, successfully assigning the minor (monomeric) peaks and obtaining the exchange rates, which cannot be achieved by ZZ-exchange alone.

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Acknowledgements

This work was supported by JSPS KAKENHI Grant Numbers JP 18H04551, JP 16K18503, JP 18K14665, and JP 19J13259.

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Authors and Affiliations

  1. Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Kyoto-Daigaku Katsura, Nishikyo-Ku, Kyoto, 615-8510, Japan

    Mayu Nishizawa, Daichi Morimoto & Kenji Sugase

  2. Department of Molecular and Cellular Physiology, Graduate School of Medicine, Kyoto University, Sakyo-ku Yoshida Konoe-cho, Kyoto, 606-8501, Japan

    Erik Walinda

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  1. Mayu Nishizawa
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  2. Erik Walinda
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  3. Daichi Morimoto
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  4. Kenji Sugase
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Correspondence to Kenji Sugase.

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Nishizawa, M., Walinda, E., Morimoto, D. et al. Pinpoint analysis of a protein in slow exchange using F1F2-selective ZZ-exchange spectroscopy: assignment and kinetic analysis. J Biomol NMR 74, 205–211 (2020). https://doi.org/10.1007/s10858-020-00309-x

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