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Optimized co-solute paramagnetic relaxation enhancement for the rapid NMR analysis of a highly fibrillogenic peptide

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Abstract

Co-solute paramagnetic relaxation enhancement (PRE) is an attractive way to speed up data acquisition in NMR spectroscopy by shortening the T 1 relaxation time of the nucleus of interest and thus the necessary recycle delay. Here, we present the rationale to utilize high-spin iron(III) as the optimal transition metal for this purpose and characterize the properties of its neutral chelate form Fe(DO3A) as a suitable PRE agent. Fe(DO3A) effectively reduces the T 1 values across the entire sequence of the intrinsically disordered protein α-synuclein with negligible impact on line width. The agent is better suited than currently used alternatives, shows no specific interaction with the polypeptide chain and, due to its high relaxivity, is effective at low concentrations and in ‘proton-less’ NMR experiments. By using Fe(DO3A) we were able to complete the backbone resonance assignment of a highly fibrillogenic peptide from α1-antitrypsin by acquiring the necessary suite of multidimensional NMR datasets in 3 h.

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Acknowledgments

Prof. Gerard Roelfes is acknowledged for access to organic synthesis facilities. Prof. Yuji Goto and Dr. Hisashi Yagi for their help with α-synuclein production. The Osaka Frontier Labs Program is gratefully acknowledged for a visiting scholarship to NAO. This work was supported by a VIDI grant from the Netherlands Organization for Scientific Research to FAAM, and by the Danish National Research Foundation (DNRF 0059).

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Author notes

  1. Takahisa Ikegami

    Present address: Graduate School of Medical Life Science, Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama, 230-0045, Japan

Authors and Affiliations

  1. Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands

    Nur Alia Oktaviani, Djurre H. de Jong, Renee Otten, Ruud M. Scheek & Frans A. A. Mulder

  2. Interdisciplinary Nanoscience Center (iNANO) and Department of Chemistry, University of Aarhus, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark

    Michael W. Risør, Niels Chr. Nielsen & Frans A. A. Mulder

  3. Institute for Protein Research, Osaka University, Suita, Japan

    Young-Ho Lee & Takahisa Ikegami

  4. Stratingh Institute for Chemistry, University of Groningen, Groningen, The Netherlands

    Rik P. Megens

  5. Interdisciplinary Nanoscience Center (iNANO) and Department of Molecular Biology and Genetics, University of Aarhus, Aarhus C, Denmark

    Jan J. Enghild

Authors
  1. Nur Alia Oktaviani
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  2. Michael W. Risør
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  11. Frans A. A. Mulder
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Corresponding author

Correspondence to Frans A. A. Mulder.

Additional information

Nur Alia Oktaviani and Michael W. Risør have equally contributed to this work.

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Oktaviani, N.A., Risør, M.W., Lee, YH. et al. Optimized co-solute paramagnetic relaxation enhancement for the rapid NMR analysis of a highly fibrillogenic peptide. J Biomol NMR 62, 129–142 (2015). https://doi.org/10.1007/s10858-015-9925-8

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  • DOI: https://doi.org/10.1007/s10858-015-9925-8

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