Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Peruzzi, Justin A., Galvez, Nina R., Kamat, Neha P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2023
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
_version_ 1785040287733645312
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
work_keys_str_mv AT peruzzijustina engineeringtransmembranesignaltransductioninsyntheticmembranesusingtwocomponentsystems
AT galvezninar engineeringtransmembranesignaltransductioninsyntheticmembranesusingtwocomponentsystems
AT kamatnehap engineeringtransmembranesignaltransductioninsyntheticmembranesusingtwocomponentsystems