European Biophysics Journal

, Volume 41, Issue 5, pp 475–482 | Cite as

Comparative analysis of the orientation of transmembrane peptides using solid-state 2H- and 15N-NMR: mobility matters

  • Stephan L. Grage
  • Erik Strandberg
  • Parvesh Wadhwani
  • Santiago Esteban-Martín
  • Jesús Salgado
  • Anne S. Ulrich
Original Paper


Many solid-state nuclear magnetic resonance (NMR) approaches for membrane proteins rely on orientation-dependent parameters, from which the alignment of peptide segments in the lipid bilayer can be calculated. Molecules embedded in liquid-crystalline membranes, such as monomeric helices, are highly mobile, leading to partial averaging of the measured NMR parameters. These dynamic effects need to be taken into account to avoid misinterpretation of NMR data. Here, we compare two common NMR approaches: 2H-NMR quadrupolar waves, and separated local field 15N–1H polarization inversion spin exchange at magic angle (PISEMA) spectra, in order to identify their strengths and drawbacks for correctly determining the orientation and mobility of α-helical transmembrane peptides. We first analyzed the model peptide WLP23 in oriented dimyristoylphosphatidylcholine (DMPC) membranes and then contrasted it with published data on GWALP23 in dilauroylphosphatidylcholine (DLPC). We only obtained consistent tilt angles from the two methods when taking dynamics into account. Interestingly, the two related peptides differ fundamentally in their mobility. Although both helices adopt the same tilt in their respective bilayers (~20°), WLP23 undergoes extensive fluctuations in its azimuthal rotation angle, whereas GWALP23 is much less dynamic. Both alternative NMR methods are suitable for characterizing orientation and dynamics, yet they can be optimally used to address different aspects. PISEMA spectra immediately reveal the presence of large-amplitude rotational fluctuations, which are not directly seen by 2H-NMR. On the other hand, PISEMA was unable to define the azimuthal rotation angle in the case of the highly dynamic WLP23, though the helix tilt could still be determined, irrespective of any dynamics parameters.


Membrane peptide orientation and dynamics WALP family peptides Geometric analysis of labeled alanines (GALA) 15N–1H PISEMA Models of peptide dynamics Selective isotope labeling 



This study was supported by the DFG Center for Functional Nanostructures CFN (E1.2) and the Spanish MICINN BFU2010-19118/BMC, financed in part by the European Regional Development Fund.

Supplementary material

249_2012_801_MOESM1_ESM.pdb (11 kb)
Structures representing the helix geometry used for the prediction of the solid-state NMR parameters are provided as pdb files in the Supplementary Materials. (PDB 10 kb)
249_2012_801_MOESM2_ESM.pdb (11 kb)
(PDB 10 kb)


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Copyright information

© European Biophysical Societies' Association 2012

Authors and Affiliations

  • Stephan L. Grage
    • 1
  • Erik Strandberg
    • 1
  • Parvesh Wadhwani
    • 1
  • Santiago Esteban-Martín
    • 2
  • Jesús Salgado
    • 3
  • Anne S. Ulrich
    • 1
  1. 1.Institute for Biological Interfaces (IBG-2) and Institute of Organic Chemistry and CFNKarlsruhe Institute of TechnologyKarlsruheGermany
  2. 2.Joint BSC-IRB Research Programme in Computational BiologyInstitute for Research in Biomedicine (IRB Barcelona), Parc Científic de BarcelonaBarcelonaSpain
  3. 3.Institute of Molecular ScienceUniversity of ValenciaPaterna (Valencia)Spain

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