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Engineering transmembrane signal transduction in synthetic membranes using two-component systems
Cells use signal transduction across their membranes to sense and respond to a wide array of chemical and physical signals. Creating synthetic systems which can harness cellular signaling modalities promises to provide a powerful platform for biosensing and therapeutic applications. As a first step...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
National Academy of Sciences
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10175788/ https://www.ncbi.nlm.nih.gov/pubmed/37126679 http://dx.doi.org/10.1073/pnas.2218610120 |
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author | Peruzzi, Justin A. Galvez, Nina R. Kamat, Neha P. |
author_facet | Peruzzi, Justin A. Galvez, Nina R. Kamat, Neha P. |
author_sort | Peruzzi, Justin A. |
collection | PubMed |
description | Cells use signal transduction across their membranes to sense and respond to a wide array of chemical and physical signals. Creating synthetic systems which can harness cellular signaling modalities promises to provide a powerful platform for biosensing and therapeutic applications. As a first step toward this goal, we investigated how bacterial two-component systems (TCSs) can be leveraged to enable transmembrane-signaling with synthetic membranes. Specifically, we demonstrate that a bacterial two-component nitrate-sensing system (NarX-NarL) can be reproduced outside of a cell using synthetic membranes and cell-free protein expression systems. We find that performance and sensitivity of the TCS can be tuned by altering the biophysical properties of the membrane in which the histidine kinase (NarX) is integrated. Through protein engineering efforts, we modify the sensing domain of NarX to generate sensors capable of detecting an array of ligands. Finally, we demonstrate that these systems can sense ligands in relevant sample environments. By leveraging membrane and protein design, this work helps reveal how transmembrane sensing can be recapitulated outside of the cell, adding to the arsenal of deployable cell-free systems primed for real world biosensing. |
format | Online Article Text |
id | pubmed-10175788 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | National Academy of Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-101757882023-11-01 Engineering transmembrane signal transduction in synthetic membranes using two-component systems Peruzzi, Justin A. Galvez, Nina R. Kamat, Neha P. Proc Natl Acad Sci U S A Biological Sciences Cells use signal transduction across their membranes to sense and respond to a wide array of chemical and physical signals. Creating synthetic systems which can harness cellular signaling modalities promises to provide a powerful platform for biosensing and therapeutic applications. As a first step toward this goal, we investigated how bacterial two-component systems (TCSs) can be leveraged to enable transmembrane-signaling with synthetic membranes. Specifically, we demonstrate that a bacterial two-component nitrate-sensing system (NarX-NarL) can be reproduced outside of a cell using synthetic membranes and cell-free protein expression systems. We find that performance and sensitivity of the TCS can be tuned by altering the biophysical properties of the membrane in which the histidine kinase (NarX) is integrated. Through protein engineering efforts, we modify the sensing domain of NarX to generate sensors capable of detecting an array of ligands. Finally, we demonstrate that these systems can sense ligands in relevant sample environments. By leveraging membrane and protein design, this work helps reveal how transmembrane sensing can be recapitulated outside of the cell, adding to the arsenal of deployable cell-free systems primed for real world biosensing. National Academy of Sciences 2023-05-01 2023-05-09 /pmc/articles/PMC10175788/ /pubmed/37126679 http://dx.doi.org/10.1073/pnas.2218610120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) . |
spellingShingle | Biological Sciences Peruzzi, Justin A. Galvez, Nina R. Kamat, Neha P. Engineering transmembrane signal transduction in synthetic membranes using two-component systems |
title | Engineering transmembrane signal transduction in synthetic membranes using two-component systems |
title_full | Engineering transmembrane signal transduction in synthetic membranes using two-component systems |
title_fullStr | Engineering transmembrane signal transduction in synthetic membranes using two-component systems |
title_full_unstemmed | Engineering transmembrane signal transduction in synthetic membranes using two-component systems |
title_short | Engineering transmembrane signal transduction in synthetic membranes using two-component systems |
title_sort | engineering transmembrane signal transduction in synthetic membranes using two-component systems |
topic | Biological Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10175788/ https://www.ncbi.nlm.nih.gov/pubmed/37126679 http://dx.doi.org/10.1073/pnas.2218610120 |
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