Letter abstract
Nature Chemical Biology 2, 314 - 318 (2006)
Published online: 7 May 2006 | doi:10.1038/nchembio793
Direct transfer of membrane proteins from bacteria to planar bilayers for rapid screening by single-channel recording
Matthew A Holden1, Lakmal Jayasinghe1,2, Oliver Daltrop1,2, Amy Mason1 & Hagan Bayley1
Although the examination of membrane proteins in planar bilayers is a powerful methodology for evaluating their pharmacology and physiological roles, introducing membrane proteins into bilayers is often a difficult process1. Here, we use a mechanical probe to transfer membrane proteins directly from Escherichia coli expression colonies to artificial lipid bilayers. In this way, single-channel electrical recordings can be made from both of the major classes of membrane proteins, 
-helix bundles and 
barrels, which are represented respectively by a K+ channel and a bacterial pore-forming toxin. Further, we examined the bicomponent toxin leukocidin (Luk), which is composed of LukF and LukS subunits. We mixed separate LukF- and LukS-expressing colonies and transferred the mixture to a planar bilayer, which generated functional Luk pores. By this means, we rapidly screened binary combinations of mutant Luk subunits for a specific function: the ability to bind a molecular adaptor. We suggest that direct transfer from cells to bilayers will be useful in several aspects of membrane proteomics and in the construction of sensor arrays.
- Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Mansfield Road, Oxford OX1 3TA, UK.
- These authors contributed equally to this work.
Correspondence to: Matthew A Holden1 e-mail: matthew.holden@chem.ox.ac.uk
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