Measuring membrane protein bond orientations in nanodiscs via residual dipolar couplings

Protein Sci. 2014 Jul;23(7):851-6. doi: 10.1002/pro.2482. Epub 2014 May 6.


Membrane proteins are involved in numerous vital biological processes. To understand membrane protein functionality, accurate structural information is required. Usually, structure determination and dynamics of membrane proteins are studied in micelles using either solution state NMR or X-ray crystallography. Even though invaluable information has been obtained by this approach, micelles are known to be far from ideal mimics of biological membranes often causing the loss or decrease of membrane protein activity. Recently, nanodiscs, which are composed of a lipid bilayer surrounded by apolipoproteins, have been introduced as a more physiological alternative than micelles for NMR investigations on membrane proteins. Here, we show that membrane protein bond orientations in nanodiscs can be obtained by measuring residual dipolar couplings (RDCs) with the outer membrane protein OmpX embedded in nanodiscs using Pf1 phage as an alignment medium. The presented collection of membrane protein RDCs in nanodiscs represents an important step toward more comprehensive structural and dynamical NMR-based investigations of membrane proteins in a natural bilayer environment.

Keywords: COCAINE; NMR; Pf1 phage; TROSY; alignment medium; membrane protein; nanodisc; residual dipolar coupling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apolipoproteins / metabolism
  • Bacterial Outer Membrane Proteins / chemistry*
  • Bacterial Outer Membrane Proteins / metabolism
  • Bacteriophage Pf1 / genetics
  • Bacteriophage Pf1 / metabolism
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / metabolism
  • Hydrogen Bonding
  • Hydrolases / chemistry*
  • Hydrolases / metabolism
  • Lipid Bilayers / chemistry*
  • Lipid Bilayers / metabolism
  • Magnetic Resonance Spectroscopy / methods*
  • Molecular Sequence Data
  • Nanostructures / chemistry*
  • Protein Conformation
  • Protein Structure, Tertiary


  • Apolipoproteins
  • Bacterial Outer Membrane Proteins
  • Escherichia coli Proteins
  • Lipid Bilayers
  • OmpX protein, E coli
  • Hydrolases

Associated data

  • PDB/2MNH