Use of reverse micelles in membrane protein structural biology |
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Authors: | Wade D Van Horn Mark E Ogilvie Peter F Flynn |
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Institution: | (1) Department of Chemistry, University of Utah, Salt Lake City, UT 84112-0850, USA;(2) Present address: Department of Biochemistry and Center for Structural Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA |
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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.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | Membrane protein Gramicidin A Encapsulation Reverse micelle PFG NMR Diffusion |
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