Nanoelectrospray based synthesis of large, transportable membranes with integrated membrane proteins

Author:

Wilm Matthias1

Affiliation:

1. University College Dublin

Abstract

Abstract

Membrane proteins tend to be difficult to study since they need to be integrated into a lipid bilayer membrane to function properly. Here I am presenting a method to synthesize a macroscopically large and freely transportable membrane with integrated membrane proteins. This is useful for studying membrane proteins and protein complexes in isolation. The method could serve as a blueprint for the production of larger quantities of functionalised membranes for integration into technical devices similar to the MinION DNA sequencer. It is possible to self-assemble larger biological membranes on solid surfaces. However, they cannot be removed from their solid support without destroying them. In transportable form, self-assembled membranes are limited to sizes of about 17 nm in nanodiscs. Here we electrospray a series of molecular layers onto the liquid surface of a buffer solution. This creates a flat, liquid environment on the surface that directs the self-assembly of the membrane. The composition of the membrane, its lipid and protein content, is experimentally controlled. We have succeeded in producing large membranes with integrated OmpG, a transmembrane pore protein. The technique appears to be compatible with the assembly of membrane based protein complexes.

Publisher

Springer Science and Business Media LLC

Reference29 articles.

1. Wilm, M. Synthesis of Extended, Self-Assembled Biological Membranes containing Membrane Proteins from Gas Phase. bioRxiv 23, 661215 (2019). Preprint at https://www.biorxiv.org/content/10.1101/661215v2

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4. Growth of large and highly ordered 2D crystals of a K⁺ channel, structural role of lipidic environment;Zorzi R;Biophysical journal,2013

5. Nanodiscs in Membrane Biochemistry and Biophysics;Denisov IG;Chemical reviews,2017

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