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An engineered protein nanopore let researchers follow a copper-catalysed click reaction molecule by molecule—and report a long-lived intermediate lasting 4.5 seconds. The 2015 experiment made the pore a tiny observation chamber for chemistry, not a miniature factory for producing compounds.
How can a nanopore act as a nanoreactor?
The researchers used α-hemolysin (αHL), a protein that forms a channel through a membrane. They engineered the channel so a chemically reactive site sat inside its pore. As reactants and reaction events affected the flow of ions through the channel, changes in ionic current provided a signal that could be monitored at the single-molecule level.
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To put a specific reactive group in the pore, Joongoo Lee and Hagan Bayley made the central transmembrane segment of the protein by solid-phase peptide synthesis. They joined it to recombinant sections on either side using native chemical ligation; the resulting protein subunits then assembled into the pore. This semisynthetic approach allowed them to place an unnatural amino acid carrying a terminal alkyne at a chosen position in the channel, rather than relying only on the side chains available in an unmodified protein. The 2015 study in PNAS describes the method and experiment.
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What click reaction did the pore observe?
In this experiment, “click reaction” refers to a copper(I)-catalysed azide–alkyne cycloaddition. An azide reacted with the alkyne attached to the pore’s unnatural amino acid, forming a triazole. The term “click chemistry” covers a family of reactions; this particular demonstration used these reactants and a copper(I) catalyst.
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The engineered pore did more than provide a site for the reaction: its ionic-current signal let the researchers follow events involving reactants inside the channel one molecule at a time. That made it possible to observe a reaction intermediate rather than infer every event only from a bulk mixture.
What did the researchers learn about the intermediate?
Lee and Bayley reported a long-lived reaction intermediate with a duration of 4.5 seconds in the study’s abstract. That is the paper’s reported observation, not a general lifetime for intermediates in click reactions. Contemporaneous Chemistry World coverage described the result as about five seconds; 4.5 seconds is the more precise figure reported by the paper.
The observation matters because it shows that a deliberately engineered protein pore can reveal a chemically interesting intermediate at single-molecule resolution. Bayley told Chemistry World, “This is a concrete result that anyone who formulates a reaction mechanism for this reaction has to take into account.”
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The nanoreactor was an engineered αHL pore used to observe a reaction, not a free-standing miniature chemical plant or a preparative synthesis device. The result demonstrates one reaction setup; it does not establish that the method can monitor every class of reaction or is routinely deployed outside research.
Later work puts the experiment in a broader sensing context. A 2021 paper on programmable nano-reactors discusses stochastic sensing applications as well as the engineering demands involved in designing and assembling suitable reactive nanopores. It is evidence of continuing research interest, not proof that the 2015 assay is commercially available or routinely used. Jia and colleagues’ 2021 Nature Communications paper provides that later context.
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