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Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers

Inclusion bodies (IBs) are typically non-functional particles of aggregated proteins. However, some proteins in fusion with amyloid-like peptides, viral coat proteins, and cellulose binding domains (CBDs) generate IB particles retaining the original functions in cells. Here, we attempted to generate...

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Autores principales: Choi, Su-Lim, Lee, Sang Jun, Yeom, Soo-Jin, Kim, Hyun Ju, Rhee, Young Ha, Jung, Heung-Chae, Lee, Seung-Goo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4045587/
https://www.ncbi.nlm.nih.gov/pubmed/24897378
http://dx.doi.org/10.1371/journal.pone.0097093
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author Choi, Su-Lim
Lee, Sang Jun
Yeom, Soo-Jin
Kim, Hyun Ju
Rhee, Young Ha
Jung, Heung-Chae
Lee, Seung-Goo
author_facet Choi, Su-Lim
Lee, Sang Jun
Yeom, Soo-Jin
Kim, Hyun Ju
Rhee, Young Ha
Jung, Heung-Chae
Lee, Seung-Goo
author_sort Choi, Su-Lim
collection PubMed
description Inclusion bodies (IBs) are typically non-functional particles of aggregated proteins. However, some proteins in fusion with amyloid-like peptides, viral coat proteins, and cellulose binding domains (CBDs) generate IB particles retaining the original functions in cells. Here, we attempted to generate CBD IBs displaying functional leucine zipper proteins (LZs) as bait for localizing cytosolic proteins in E. coli. When a red fluorescent protein was tested as a target protein, microscopic observations showed that the IBs red-fluoresced strongly. When different LZ pairs with K(D)s of 8–1,000 µM were tested as the bait and prey, the localization of the red fluorescence appeared to change following the affinities between the LZs, as observed by fluorescence imaging and flow cytometry. This result proposed that LZ-tagged CBD IBs can be applied as an in vivo matrix to entrap cytosolic proteins in E. coli while maintaining their original activities. In addition, easy detection of localization to IBs provides a unique platform for the engineering and analyses of protein-protein interactions in E. coli.
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spelling pubmed-40455872014-06-09 Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers Choi, Su-Lim Lee, Sang Jun Yeom, Soo-Jin Kim, Hyun Ju Rhee, Young Ha Jung, Heung-Chae Lee, Seung-Goo PLoS One Research Article Inclusion bodies (IBs) are typically non-functional particles of aggregated proteins. However, some proteins in fusion with amyloid-like peptides, viral coat proteins, and cellulose binding domains (CBDs) generate IB particles retaining the original functions in cells. Here, we attempted to generate CBD IBs displaying functional leucine zipper proteins (LZs) as bait for localizing cytosolic proteins in E. coli. When a red fluorescent protein was tested as a target protein, microscopic observations showed that the IBs red-fluoresced strongly. When different LZ pairs with K(D)s of 8–1,000 µM were tested as the bait and prey, the localization of the red fluorescence appeared to change following the affinities between the LZs, as observed by fluorescence imaging and flow cytometry. This result proposed that LZ-tagged CBD IBs can be applied as an in vivo matrix to entrap cytosolic proteins in E. coli while maintaining their original activities. In addition, easy detection of localization to IBs provides a unique platform for the engineering and analyses of protein-protein interactions in E. coli. Public Library of Science 2014-06-04 /pmc/articles/PMC4045587/ /pubmed/24897378 http://dx.doi.org/10.1371/journal.pone.0097093 Text en © 2014 Choi et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Choi, Su-Lim
Lee, Sang Jun
Yeom, Soo-Jin
Kim, Hyun Ju
Rhee, Young Ha
Jung, Heung-Chae
Lee, Seung-Goo
Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers
title Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers
title_full Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers
title_fullStr Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers
title_full_unstemmed Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers
title_short Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers
title_sort controlled localization of functionally active proteins to inclusion bodies using leucine zippers
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4045587/
https://www.ncbi.nlm.nih.gov/pubmed/24897378
http://dx.doi.org/10.1371/journal.pone.0097093
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