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Tunable paramagnetic relaxation enhancements by [Gd(DPA)3]3− for protein structure analysis

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Abstract

Paramagnetic relaxation enhancements (PRE) present a powerful source of structural information in nuclear magnetic resonance (NMR) studies of proteins and protein–ligand complexes. In contrast to conventional PRE reagents that are covalently attached to the protein, the complex between gadolinium and three dipicolinic acid (DPA) molecules, [Gd(DPA)3]3−, can bind to proteins in a non-covalent yet site-specific manner. This offers straightforward access to PREs that can be scaled by using different ratios of [Gd(DPA)3]3− to protein, allowing quantitative distance measurements for nuclear spins within about 15 Å of the Gd3+ ion. Such data accurately define the metal position relative to the protein, greatly enhancing the interpretation of pseudocontact shifts induced by [Ln(DPA)3]3− complexes of paramagnetic lanthanide (Ln3+) ions other than gadolinium. As an example we studied the quaternary structure of the homodimeric GCN4 leucine zipper.

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Acknowledgments

Financial support by the Australian Research Council is gratefully acknowledged.

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Authors and Affiliations

  1. Research School of Chemistry, Australian National University, Canberra, ACT, 0200, Australia

    Hiromasa Yagi, Karin V. Loscha, Xun-Cheng Su & Gottfried Otting

  2. School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, QLD, 4072, Australia

    Mitchell Stanton-Cook & Thomas Huber

Authors
  1. Hiromasa Yagi
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  2. Karin V. Loscha
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  3. Xun-Cheng Su
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  4. Mitchell Stanton-Cook
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  5. Thomas Huber
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  6. Gottfried Otting
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Correspondence to Gottfried Otting.

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Yagi, H., Loscha, K.V., Su, XC. et al. Tunable paramagnetic relaxation enhancements by [Gd(DPA)3]3− for protein structure analysis. J Biomol NMR 47, 143–153 (2010). https://doi.org/10.1007/s10858-010-9416-x

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  • DOI: https://doi.org/10.1007/s10858-010-9416-x

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