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A real-time analysis of protein transport via the twin arginine translocation pathway in response to different components of the protonmotive force

The twin arginine translocation (Tat) pathway transports folded protein across the cytoplasmic membrane in bacteria, archaea, and across the thylakoid membrane in plants as well as the inner membrane in some mitochondria. In plant chloroplasts, the Tat pathway utilizes the protonmotive force (PMF) t...

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Autores principales: Zhou, Wenjie, Hao, Binhan, Bricker, Terry M., Theg, Steven M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641609/
https://www.ncbi.nlm.nih.gov/pubmed/37742925
http://dx.doi.org/10.1016/j.jbc.2023.105286
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author Zhou, Wenjie
Hao, Binhan
Bricker, Terry M.
Theg, Steven M.
author_facet Zhou, Wenjie
Hao, Binhan
Bricker, Terry M.
Theg, Steven M.
author_sort Zhou, Wenjie
collection PubMed
description The twin arginine translocation (Tat) pathway transports folded protein across the cytoplasmic membrane in bacteria, archaea, and across the thylakoid membrane in plants as well as the inner membrane in some mitochondria. In plant chloroplasts, the Tat pathway utilizes the protonmotive force (PMF) to drive protein translocation. However, in bacteria, it has been shown that Tat transport depends only on the transmembrane electrical potential (Δψ) component of PMF in vitro. To investigate the comprehensive PMF requirement in Escherichia coli, we have developed the first real-time assay to monitor Tat transport utilizing the NanoLuc Binary Technology in E. coli spheroplasts. This luminescence assay allows for continuous monitoring of Tat transport with high-resolution, making it possible to observe subtle changes in transport in response to different treatments. By applying the NanoLuc assay, we report that, under acidic conditions (pH = 6.3), ΔpH, in addition to Δψ, contributes energetically to Tat transport in vivo in E. coli spheroplasts. These results provide novel insight into the mechanism of energy utilization by the Tat pathway.
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spelling pubmed-106416092023-11-14 A real-time analysis of protein transport via the twin arginine translocation pathway in response to different components of the protonmotive force Zhou, Wenjie Hao, Binhan Bricker, Terry M. Theg, Steven M. J Biol Chem Research Article Collection: Bioenergetics The twin arginine translocation (Tat) pathway transports folded protein across the cytoplasmic membrane in bacteria, archaea, and across the thylakoid membrane in plants as well as the inner membrane in some mitochondria. In plant chloroplasts, the Tat pathway utilizes the protonmotive force (PMF) to drive protein translocation. However, in bacteria, it has been shown that Tat transport depends only on the transmembrane electrical potential (Δψ) component of PMF in vitro. To investigate the comprehensive PMF requirement in Escherichia coli, we have developed the first real-time assay to monitor Tat transport utilizing the NanoLuc Binary Technology in E. coli spheroplasts. This luminescence assay allows for continuous monitoring of Tat transport with high-resolution, making it possible to observe subtle changes in transport in response to different treatments. By applying the NanoLuc assay, we report that, under acidic conditions (pH = 6.3), ΔpH, in addition to Δψ, contributes energetically to Tat transport in vivo in E. coli spheroplasts. These results provide novel insight into the mechanism of energy utilization by the Tat pathway. American Society for Biochemistry and Molecular Biology 2023-09-22 /pmc/articles/PMC10641609/ /pubmed/37742925 http://dx.doi.org/10.1016/j.jbc.2023.105286 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article Collection: Bioenergetics
Zhou, Wenjie
Hao, Binhan
Bricker, Terry M.
Theg, Steven M.
A real-time analysis of protein transport via the twin arginine translocation pathway in response to different components of the protonmotive force
title A real-time analysis of protein transport via the twin arginine translocation pathway in response to different components of the protonmotive force
title_full A real-time analysis of protein transport via the twin arginine translocation pathway in response to different components of the protonmotive force
title_fullStr A real-time analysis of protein transport via the twin arginine translocation pathway in response to different components of the protonmotive force
title_full_unstemmed A real-time analysis of protein transport via the twin arginine translocation pathway in response to different components of the protonmotive force
title_short A real-time analysis of protein transport via the twin arginine translocation pathway in response to different components of the protonmotive force
title_sort real-time analysis of protein transport via the twin arginine translocation pathway in response to different components of the protonmotive force
topic Research Article Collection: Bioenergetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641609/
https://www.ncbi.nlm.nih.gov/pubmed/37742925
http://dx.doi.org/10.1016/j.jbc.2023.105286
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