Journal of Biomolecular NMR

, Volume 47, Issue 2, pp 143–153

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

  • Hiromasa Yagi
  • Karin V. Loscha
  • Xun-Cheng Su
  • Mitchell Stanton-Cook
  • Thomas Huber
  • Gottfried Otting
Article

DOI: 10.1007/s10858-010-9416-x

Cite this article as:
Yagi, H., Loscha, K.V., Su, XC. et al. J Biomol NMR (2010) 47: 143. doi:10.1007/s10858-010-9416-x

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.

Keywords

Lanthanide tagsLeucine zipperNMR spectroscopyParamagnetic relaxation enhancementsPseudocontact shifts

Supplementary material

10858_2010_9416_MOESM1_ESM.pdf (1.1 mb)
Supplementary material 1 (PDF 1169 kb)

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Hiromasa Yagi
    • 1
  • Karin V. Loscha
    • 1
  • Xun-Cheng Su
    • 1
  • Mitchell Stanton-Cook
    • 2
  • Thomas Huber
    • 2
  • Gottfried Otting
    • 1
  1. 1.Research School of ChemistryAustralian National UniversityCanberraAustralia
  2. 2.School of Chemistry and Molecular BiosciencesUniversity of QueenslandBrisbaneAustralia