Protease-based synthetic sensing and signal amplification

Stein, Viktor and Alexandrov, Kirill (2014) Protease-based synthetic sensing and signal amplification. Proceedings of the National Academy of Sciences of the United States of America, 111 45: 15934-15939. doi:10.1073/pnas.1405220111

Author Stein, Viktor
Alexandrov, Kirill
Title Protease-based synthetic sensing and signal amplification
Journal name Proceedings of the National Academy of Sciences of the United States of America   Check publisher's open access policy
ISSN 1091-6490
Publication date 2014-11-11
Sub-type Article (original research)
DOI 10.1073/pnas.1405220111
Open Access Status
Volume 111
Issue 45
Start page 15934
End page 15939
Total pages 6
Place of publication Washington, DC, United States
Publisher National Academy of Sciences
Collection year 2015
Language eng
Abstract The bottom-up design of protein-based signaling networks is a key goal of synthetic biology; yet, it remains elusive due to our inability to tailor-make signal transducers and receptors that can be readily compiled into defined signaling networks. Here, we report a generic approach for the construction of protein-based molecular switches based on artficially autoinhibited proteases. Using structure-guided design and directed protein evolution, we created signal transducers based on artificially autoinhibited proteases that can be activated following site-specific proteolysis and also demonstrate the modular design of an allosterically regulated protease receptor following recombination with an affinity clamp peptide receptor. Notably, the receptor's mode of action can be varied from >5-fold switch-OFF to >30-fold switch-ON solely by changing the length of the connecting linkers, demonstrating a high functional plasticity not previously observed in naturally occurring receptor systems. We also create an integrated signaling circuit based on two orthogonal autoinhibited protease units that can propagate and amplify molecular queues generated by the protease receptor. Finally, we present a generic two-component receptor architecture based on proximity-based activation of two autoinhibited proteases. Overall, the approach allows the design of protease-based signaling networks that, in principle, can be connected to any biological process.
Keyword Proteases
Protein engineering
Protein switches
Synthetic biology
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

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