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In Vivo Incorporation of Photoproteins into GroEL Chaperonin Retaining Major Structural and Functional Properties
The incorporation of photoproteins into proteins of interest allows the study of either their localization or intermolecular interactions in the cell. Here we demonstrate the possibility of in vivo incorporating the photoprotein Aequorea victoria enhanced green fluorescent protein (EGFP) or Gaussia...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9965216/ https://www.ncbi.nlm.nih.gov/pubmed/36838891 http://dx.doi.org/10.3390/molecules28041901 |
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author | Marchenkov, Victor Ivashina, Tanya Marchenko, Natalia Ryabova, Natalya Selivanova, Olga Timchenko, Alexander Kihara, Hiroshi Ksenzenko, Vladimir Semisotnov, Gennady |
author_facet | Marchenkov, Victor Ivashina, Tanya Marchenko, Natalia Ryabova, Natalya Selivanova, Olga Timchenko, Alexander Kihara, Hiroshi Ksenzenko, Vladimir Semisotnov, Gennady |
author_sort | Marchenkov, Victor |
collection | PubMed |
description | The incorporation of photoproteins into proteins of interest allows the study of either their localization or intermolecular interactions in the cell. Here we demonstrate the possibility of in vivo incorporating the photoprotein Aequorea victoria enhanced green fluorescent protein (EGFP) or Gaussia princeps luciferase (GLuc) into the tetradecameric quaternary structure of GroEL chaperonin and describe some physicochemical properties of the labeled chaperonin. Using size-exclusion and affinity chromatography, electrophoresis, fluorescent and electron transmission microscopy (ETM), small-angle X-ray scattering (SAXS), and bioluminescence resonance energy transfer (BRET), we show the following: (i) The GroEL(14)-EGFP is evenly distributed within normally divided E. coli cells, while gigantic undivided cells are characterized by the uneven distribution of the labeled GroEL(14) which is mainly localized close to the cellular periplasm; (ii) EGFP and likely GLuc are located within the inner cavity of one of the two GroEL chaperonin rings and do not essentially influence the protein oligomeric structure; (iii) GroEL(14) containing either EGFP or GLuc is capable of interacting with non-native proteins and the cochaperonin GroES. |
format | Online Article Text |
id | pubmed-9965216 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-99652162023-02-26 In Vivo Incorporation of Photoproteins into GroEL Chaperonin Retaining Major Structural and Functional Properties Marchenkov, Victor Ivashina, Tanya Marchenko, Natalia Ryabova, Natalya Selivanova, Olga Timchenko, Alexander Kihara, Hiroshi Ksenzenko, Vladimir Semisotnov, Gennady Molecules Article The incorporation of photoproteins into proteins of interest allows the study of either their localization or intermolecular interactions in the cell. Here we demonstrate the possibility of in vivo incorporating the photoprotein Aequorea victoria enhanced green fluorescent protein (EGFP) or Gaussia princeps luciferase (GLuc) into the tetradecameric quaternary structure of GroEL chaperonin and describe some physicochemical properties of the labeled chaperonin. Using size-exclusion and affinity chromatography, electrophoresis, fluorescent and electron transmission microscopy (ETM), small-angle X-ray scattering (SAXS), and bioluminescence resonance energy transfer (BRET), we show the following: (i) The GroEL(14)-EGFP is evenly distributed within normally divided E. coli cells, while gigantic undivided cells are characterized by the uneven distribution of the labeled GroEL(14) which is mainly localized close to the cellular periplasm; (ii) EGFP and likely GLuc are located within the inner cavity of one of the two GroEL chaperonin rings and do not essentially influence the protein oligomeric structure; (iii) GroEL(14) containing either EGFP or GLuc is capable of interacting with non-native proteins and the cochaperonin GroES. MDPI 2023-02-16 /pmc/articles/PMC9965216/ /pubmed/36838891 http://dx.doi.org/10.3390/molecules28041901 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Marchenkov, Victor Ivashina, Tanya Marchenko, Natalia Ryabova, Natalya Selivanova, Olga Timchenko, Alexander Kihara, Hiroshi Ksenzenko, Vladimir Semisotnov, Gennady In Vivo Incorporation of Photoproteins into GroEL Chaperonin Retaining Major Structural and Functional Properties |
title | In Vivo Incorporation of Photoproteins into GroEL Chaperonin Retaining Major Structural and Functional Properties |
title_full | In Vivo Incorporation of Photoproteins into GroEL Chaperonin Retaining Major Structural and Functional Properties |
title_fullStr | In Vivo Incorporation of Photoproteins into GroEL Chaperonin Retaining Major Structural and Functional Properties |
title_full_unstemmed | In Vivo Incorporation of Photoproteins into GroEL Chaperonin Retaining Major Structural and Functional Properties |
title_short | In Vivo Incorporation of Photoproteins into GroEL Chaperonin Retaining Major Structural and Functional Properties |
title_sort | in vivo incorporation of photoproteins into groel chaperonin retaining major structural and functional properties |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9965216/ https://www.ncbi.nlm.nih.gov/pubmed/36838891 http://dx.doi.org/10.3390/molecules28041901 |
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