Micro-coil NMR to monitor optimization of the reconstitution conditions for the integral membrane protein OmpW in detergent micelles

J Biomol NMR. 2012 Oct;54(2):129-33. doi: 10.1007/s10858-012-9658-x. Epub 2012 Aug 14.

Abstract

Optimization of aqueous solutions of the integral membrane protein (IMP) OmpW for NMR structure determination has been monitored with micro-coil NMR, which enables the acquisition of NMR spectra using only micrograms of protein and detergent. The detergent 30-Fos (2-undecylphosphocholine) was found to yield the best 2D [(15)N, (1)H]-TROSY correlation NMR spectra of [(2)H, (15)N]-labeled OmpW. For the OmpW structure determination we then optimized the 30-Fos concentration, the sample temperature and long-time stability, and the deuteration level of the protein. Some emerging guidelines for reconstitution of β-barrel integral membrane proteins in structural biology are discussed.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Outer Membrane Proteins / chemistry*
  • Detergents / chemistry*
  • Deuterium
  • Escherichia coli Proteins / chemistry*
  • Micelles
  • Phosphorylcholine / analogs & derivatives*
  • Phosphorylcholine / chemistry
  • Protein Conformation
  • Protein Stability

Substances

  • 2-undecylphosphocholine
  • Bacterial Outer Membrane Proteins
  • Detergents
  • Escherichia coli Proteins
  • Micelles
  • ompW protein, E coli
  • Phosphorylcholine
  • Deuterium