Residual dipolar couplings: are multiple independent alignments always possible?

Victoria A. Higman, Jonathan Boyd, Lorna J. Smith*, Christina Redfield

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

24 Citations (Scopus)

Abstract

RDCs for the 14 kDa protein hen egg-white lysozyme (HEWL) have been measured in eight different alignment media. The elongated shape and strongly positively charged surface of HEWL appear to limit the protein to four main alignment orientations. Furthermore, low levels of alignment and the protein's interaction with some alignment media increases the experimental error. Together with heterogeneity across the alignment media arising from constraints on temperature, pH and ionic strength for some alignment media, these data are suitable for structure refinement, but not the extraction of dynamic parameters. For an analysis of protein dynamics the data must be obtained with very low errors in at least three or five independent alignment media (depending on the method used) and so far, such data have only been reported for three small 6-8 kDa proteins with identical folds: ubiquitin, GB1 and GB3. Our results suggest that HEWL is likely to be representative of many other medium to large sized proteins commonly studied by solution NMR. Comparisons with over 60 high-resolution crystal structures of HEWL reveal that the highest resolution structures are not necessarily always the best models for the protein structure in solution.

Original languageEnglish
Pages (from-to)53-60
Number of pages8
JournalJournal of Biomolecular NMR
Volume49
Issue number1
DOIs
Publication statusPublished - Jan 2011

Keywords

  • Hen egg-white lysozyme
  • HEN LYSOZYME
  • Dynamics
  • CONSERVATIVE MUTAGENESIS
  • Residual dipolar couplings
  • MEDIA
  • CHEMICAL-SHIFTS
  • EGG-WHITE LYSOZYME
  • PROTEIN BACKBONE
  • DYNAMIC INTERPRETATION
  • N-15 NMR RELAXATION
  • Protein alignment
  • Ubiquitin
  • LIQUID-CRYSTALLINE MEDIUM
  • MODEL-FREE ANALYSIS

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