Journal of Biomolecular NMR

, Volume 43, Issue 4, pp 219–227

Selective backbone labelling of ILV methyl labelled proteins

  • Nathalie Sibille
  • Xavier Hanoulle
  • Fanny Bonachera
  • Dries Verdegem
  • Isabelle Landrieu
  • Jean-Michel Wieruszeski
  • Guy Lippens
Article

Abstract

Adding the 13C labelled 2-keto-isovalerate and 2-oxobutanoate precursors to a minimal medium composed of 12C labelled glucose instead of the commonly used (2D, 13C) glucose leads not only to the 13C labelling of (I, L, V) methyls but also to the selective 13C labelling of the backbone Cα and CO carbons of the Ile and Val residues. As a result, the backbone (1H, 15N) correlations of the Ile and Val residues and their next neighbours in the (i + 1) position can be selectively identified in HN(CA) and HN(CO) planes. The availability of a selective HSQC spectrum corresponding to the sole amide resonances of the Ile and Val residues allows connecting them to their corresponding methyls by the intra-residue NOE effect, and should therefore be applicable to larger systems.

Keywords

NMR spectroscopy Assignment Selective labelling Methyl trosy 

Abbreviations

TauF1

Residues 163-441 of the Tau protein

HMQC

Heteronuclear multiple quantum coherence

HSQC

Heteronuclear single quantum coherence

NMR

Nuclear magnetic resonance

NOESY

Nuclear Overhauser Effect spectroscopy

TOCSY

Total correlation spectroscopy

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Nathalie Sibille
    • 1
  • Xavier Hanoulle
    • 1
  • Fanny Bonachera
    • 1
  • Dries Verdegem
    • 1
  • Isabelle Landrieu
    • 1
  • Jean-Michel Wieruszeski
    • 1
  • Guy Lippens
    • 1
  1. 1.CNRS UMR 8576 – Unité de Glycobiologie Structurale et FonctionnelleUniversité des Sciences et Technologies de Lille 1Villeneuve d’Ascq CedexFrance

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