Stochastic sensing of organic analytes by a pore-forming protein containing a molecular adapter

Li-Qun Gu, Orit Braha (), Sean Conlan, Stephen Cheley and Hagan Bayley ()
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Li-Qun Gu: Texas A&M University Health Science Center
Orit Braha: Texas A&M University Health Science Center
Sean Conlan: Texas A&M University Health Science Center
Stephen Cheley: Texas A&M University Health Science Center
Hagan Bayley: Texas A&M University

Nature, 1999, vol. 398, issue 6729, 686-690

Abstract: Abstract The detection of organic molecules is important in many areas, including medicine, environmental monitoring and defence1,5. Stochastic sensing is an approach that relies on the observation of individual binding events between analyte molecules and a single receptor6. Engineered transmembrane protein pores are promising sensor elements for stochastic detection6, and in their simplest manifestation they produce a fluctuating binary (‘on/off’) response in the transmembrane electrical current. The frequency of occurrence of the fluctuations reveals the concentration of the analyte, and its identity can be deduced from the characteristic magnitude and/or duration of the fluctuations. Genetically engineered versions of the bacterial pore-forming protein α-haemolysin have been used to identify and quantify divalent metal ions in solution6. But it is not immediately obvious how versatile binding sites for organic ligands might be obtained by engineering of the pore structure. Here we show that stochastic sensing of organic molecules can be procured from α-haemolysin by equipping the channel with an internal, non-covalently bound molecular ‘adapter’ which mediates channel blocking by the analyte. We use cyclodextrins as the adapters because these fit comfortably inside the pore and present a hydrophobic cavity suitable for binding a variety of organic analytes. Moreover, a single sensing element of this sort can be used to analyse a mixture of organic molecules with different binding characteristics. We envisage the use of other adapters, so that the pore could be ‘programmed’ for a range of sensing functions.

Date: 1999
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DOI: 10.1038/19491

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