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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2023
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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. |
format | Online Article Text |
id | pubmed-10641609 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
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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