Connecting Artificial Proteolytic and Electrochemical Signaling Systems with Caged Messenger Peptides
| Autor: | Selvakumar Edwardraja, Artem Melman, Paolo Bollella, Jason Whitfield, Patricia M. Walden, Claudia E. Vickers, Evgeny Katz, Zhong Guo, Kirill Alexandrov |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Calmodulin
medicine.medical_treatment Proteolysis Allosteric regulation Bioengineering Peptide Biosensing Techniques 010402 general chemistry 01 natural sciences 03 medical and health sciences medicine Instrumentation 030304 developmental biology Fluid Flow and Transfer Processes chemistry.chemical_classification 0303 health sciences Protease biology medicine.diagnostic_test Effector Process Chemistry and Technology In vitro 0104 chemical sciences chemistry Biophysics biology.protein Biosensor Peptide Hydrolases Signal Transduction |
| Zdroj: | ACS Sensors. 6:3596-3603 |
| ISSN: | 2379-3694 |
| Popis: | Enzymatic polypeptide proteolysis is a widespread and powerful biological control mechanism. Over the last few years, substantial progress has been made in creating artificial proteolytic systems where an input of choice modulates the protease activity and thereby the activity of its substrates. However, all proteolytic systems developed so far have relied on the direct proteolytic cleavage of their effectors. Here, we propose a new concept where protease biosensors with a tunable input uncage a signaling peptide, which can then transmit a signal to an allosteric protein reporter. We demonstrate that both the cage and the regulatory domain of the reporter can be constructed from the same peptide-binding domain, such as calmodulin. To demonstrate this concept, we constructed a proteolytic rapamycin biosensor and demonstrated its quantitative actuation on fluorescent, luminescent, and electrochemical reporters. Using the latter, we constructed sensitive bioelectrodes that detect the messenger peptide release and quantitatively convert the recognition event into electric current. We discuss the application of such systems for the construction of in vitro sensory arrays and in vivo signaling circuits. |
| Databáze: | OpenAIRE |
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