Use of reverse micelles in membrane protein structural biology

Wade D. van Horn, Mark E. Ogilvie, Peter F. Flynn

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Membrane protein structural biology is a rapidly developing field with fundamental importance for elucidating key biological and biophysical processes including signal transduction, intercellular communication, and cellular transport. In addition to the intrinsic interest in this area of research, structural studies of membrane proteins have direct significance on the development of therapeutics that impact human health in diverse and important ways. In this article we demonstrate the potential of investigating the structure of membrane proteins using the reverse micelle forming surfactant dioctyl sulfosuccinate (AOT) in application to the prototypical model ion channel gramicidin A. Reverse micelles are surfactant based nanoparticles which have been employed to investigate fundamental physical properties of biomolecules. The results of this solution NMR based study indicate that the AOT reverse micelle system is capable of refolding and stabilizing relatively high concentrations of the native conformation of gramicidin A. Importantly, pulsed-field-gradient NMR diffusion and NOESY experiments reveal stable gramicidin A homodimer interactions that bridge reverse micelle particles. The spectroscopic benefit of reverse micelle-membrane protein solubilization is also explored, and significant enhancement over commonly used micelle based mimetic systems is demonstrated. These results establish the effectiveness of reverse micelle based studies of membrane proteins, and illustrate that membrane proteins solubilized by reverse micelles are compatible with high resolution solution NMR techniques.

Original languageEnglish (US)
Pages (from-to)203-211
Number of pages9
JournalJournal of Biomolecular NMR
Volume40
Issue number3
DOIs
StatePublished - Mar 1 2008
Externally publishedYes

Keywords

  • Encapsulation
  • Gramicidin A
  • Membrane protein
  • PFG NMR Diffusion
  • Reverse micelle

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

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