Simultaneous measurement of ¹H-¹⁵N and methyl ¹Hm-¹³Cm residual dipolar couplings in large proteins

J Biomol NMR. 2011 Sep;51(1-2):191-8. doi: 10.1007/s10858-011-9553-x. Epub 2011 Sep 27.

Abstract

A two-dimensional TROSY-based SIM-(13)C(m)-(1)H(m)/(1)H-(15)N NMR experiment for simultaneous measurements of methyl (1) D (CH) and backbone amide (1) D (NH) residual dipolar couplings (RDC) in {U-[(15)N,(2)H]; Ileδ1-[(13)CH(3)]; Leu,Val-[(13)CH(3)/(12)CD(3)]}-labeled samples of large proteins is described. Significant variation in the alignment tensor of the 82-kDa enzyme Malate synthase G is observed as a function of only slight changes in experimental conditions. The SIM-(13)C(m)-(1)H(m)/(1)H-(15)N data sets provide convenient means of establishing the alignment tensor characteristics via the measurement of (1) D (NH) RDCs in the same protein sample.

MeSH terms

  • Carbon Isotopes
  • Leucine / genetics
  • Malate Synthase / chemistry
  • Models, Molecular
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Protein Conformation
  • Proteins / chemistry*
  • Proteins / genetics
  • Valine / genetics

Substances

  • Carbon Isotopes
  • Proteins
  • Malate Synthase
  • Leucine
  • Valine