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bioRxiv · 10.1101/678656

Engineering an Enzyme for Direct Electrical Monitoring of Activity

Zhang, B.·Deng, H.·Mukherjee, S.·Song, W.·Wang, X.·Lindsay, S.
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Abstract

Proteins have been shown to be electrically-conductive if tethered to an electrode by means of a specific binding agent, allowing single molecules to be wired into an electrical sensing circuit. This development opens the possibility of exploiting the remarkable chemical versatility of enzymes as sensors, detectors and sequencing devices. We have engineered contact points into a {Phi}29 polymerase by introducing biotinylatable peptide sequences. The modified enzyme was bound to electrodes functionalized with streptavidin. {Phi}29 connected by one biotinylated contact and a second non-specific contact showed rapid small fluctuations in current when activated. Signals were greatly enhanced with two specific contacts. Features in the distributions of DC conductance increased by a factor 2 or more over the open-to closed conformational transition of the polymerase. Polymerase activity is manifested by rapid (millisecond) large (25% of background) current fluctuations imposed on the DC conductance.

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Zhang, B., Deng, H., Mukherjee, S., Song, W., Wang, X., Lindsay, S.. 2019-06-21. Engineering an Enzyme for Direct Electrical Monitoring of Activity. https://doi.org/10.1101/678656

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