3 years ago

Ultrasensitive Scaffold-Dependent Protease Sensors with Large Dynamic Range

Ultrasensitive Scaffold-Dependent Protease Sensors with Large Dynamic Range
Masuda Nabi, Kirill Alexandrov, Viktor Stein
The rational construction of synthetic protein switches with predefined input–output parameters constitutes a key goal of synthetic biology with many potential applications ranging from metabolic engineering to diagnostics. Yet, generally applicable strategies to construct tailor-engineered protein switches have so far remained elusive. Here, we use SpyTag/SpyCatcher-mediated protein ligation to engineer modularly organized, scaffold-dependent protease sensors that exploit a combination of affinity targeting and protease-inducible protein–protein interactions. We use this architecture to create a suite of integrated signal sensing and amplification circuits that can detect the activity of α-thrombin and prostate specific antigen with a dynamic range covering 5 orders of magnitude. We determine the key design features critical for signal transmission between protease-based sensors, transducers, and actuators.

Publisher URL: http://dx.doi.org/10.1021/acssynbio.6b00370

DOI: 10.1021/acssynbio.6b00370

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