Translational diffusion measurements by microcoil NMR in aqueous solutions of the Fos-10 detergent-solubilized membrane protein OmpX

J Phys Chem B. 2012 Jun 14;116(23):6775-80. doi: 10.1021/jp212401w. Epub 2012 Feb 29.

Abstract

Aqueous solutions of the detergent Fos-10 (n-decylphosphocholine) without and with addition of the integral membrane protein (IMP) OmpX (outer membrane protein X) have been characterized using pulsed field gradient-stimulated echo (PFG-STE) NMR experiments for measurements of translational diffusion coefficients. Effective diffusion coefficients for Fos-10 micelles in the absence of OmpX were obtained by observation of NMR signals from 10-bromodecan-1-ol that had been inserted into the micelles, and in the presence of OmpX by NMR observation of the protein. It is thus shown that solutions of Fos-10-reconstituted OmpX can be quantitatively described as a mixture of Fos-10 monomers, uniform Fos-10 micelles, and uniform OmpX-containing Fos-10 micelles, with Fos-10 monomers in fast exchange between the pools of these three species. This result establishes an avenue for efficient determination of the effective translational diffusion coefficients of IMP-containing detergent micelles based on observation of the intense detergent NMR signals, which is also applicable with unlabeled IMPs. This monitoring of the species present in a given IMP solution contributes to improved guidelines for rational selection of detergent and buffer conditions in structural studies of integral membrane proteins.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacterial Outer Membrane Proteins / chemistry*
  • Detergents / chemistry*
  • Diffusion
  • Escherichia coli Proteins / chemistry*
  • Hydrolases / chemistry*
  • Models, Molecular
  • Nuclear Magnetic Resonance, Biomolecular*
  • Phosphorylcholine / analogs & derivatives*
  • Phosphorylcholine / chemistry
  • Solubility
  • Solutions
  • Water / chemistry

Substances

  • Bacterial Outer Membrane Proteins
  • Detergents
  • Escherichia coli Proteins
  • Solutions
  • Water
  • Phosphorylcholine
  • OmpX protein, E coli
  • N-decylphosphorylcholine
  • Hydrolases