Journal of Biomolecular NMR

, Volume 54, Issue 2, pp 129–133 | Cite as

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

  • Pawel Stanczak
  • Qinghai Zhang
  • Reto Horst
  • Pedro Serrano
  • Kurt Wüthrich
Communication

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 [15N, 1H]-TROSY correlation NMR spectra of [2H, 15N]-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.

Keywords

Structural biology Integral membrane proteins Solubilization in detergent micelles E. coli outer membrane protein W 

Notes

Acknowledgments

This work was supported by the Joint Center for Innovative Membrane Protein Technologies (JCIMPT-complexes, NIH Roadmap Grant P50 GM073197). K.W. is the Cecil H. and Ida M. Green Professor of Structural Biology at The Scripps Research Institute.

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Pawel Stanczak
    • 1
  • Qinghai Zhang
    • 1
  • Reto Horst
    • 1
  • Pedro Serrano
    • 1
  • Kurt Wüthrich
    • 1
    • 2
  1. 1.Department of Molecular BiologyThe Scripps Research InstituteLa JollaUSA
  2. 2.Skaggs Institute for Chemical BiologyThe Scripps Research InstituteLa JollaUSA

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