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Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli

Bioorthogonal reactions, especially the Cu(I)-catalyzed azide-alkyne cycloaddition, have revolutionized our ability to label and manipulate biomolecules under living conditions. The cytotoxicity of Cu(I) ions, however, has hindered the application of this reaction in the internal space of living cel...

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Detalles Bibliográficos
Autores principales: Yang, Maiyun, Jalloh, Abubakar S., Wei, Wei, Zhao, Jing, Wu, Peng, Chen, Peng R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4174402/
https://www.ncbi.nlm.nih.gov/pubmed/25236616
http://dx.doi.org/10.1038/ncomms5981
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author Yang, Maiyun
Jalloh, Abubakar S.
Wei, Wei
Zhao, Jing
Wu, Peng
Chen, Peng R.
author_facet Yang, Maiyun
Jalloh, Abubakar S.
Wei, Wei
Zhao, Jing
Wu, Peng
Chen, Peng R.
author_sort Yang, Maiyun
collection PubMed
description Bioorthogonal reactions, especially the Cu(I)-catalyzed azide-alkyne cycloaddition, have revolutionized our ability to label and manipulate biomolecules under living conditions. The cytotoxicity of Cu(I) ions, however, has hindered the application of this reaction in the internal space of living cells. By systematically surveying a panel of Cu(I)-stabilizing ligands in promoting protein labeling within the cytoplasm of E. coli, here we identify a highly efficient and biocompatible catalyst for intracellular modification of proteins by azide-alkyne cycloaddition. This reaction permits us to conjugate an environment-sensitive fluorophore site-specifically onto HdeA, an acid-stress chaperone that adopts pH-dependent conformational changes, in both the periplasm and cytoplasm of E. coli. The resulting protein-fluorophore hybrid pH indicators enable compartment-specific pH measurement to determine the pH gradient across the E. coli cytoplasmic membrane. This construct also allows the measurement of E. coli transmembrane potential, and the determination of the proton motive force across its inner membrane under normal and acid-stress conditions.
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spelling pubmed-41744022015-03-19 Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli Yang, Maiyun Jalloh, Abubakar S. Wei, Wei Zhao, Jing Wu, Peng Chen, Peng R. Nat Commun Article Bioorthogonal reactions, especially the Cu(I)-catalyzed azide-alkyne cycloaddition, have revolutionized our ability to label and manipulate biomolecules under living conditions. The cytotoxicity of Cu(I) ions, however, has hindered the application of this reaction in the internal space of living cells. By systematically surveying a panel of Cu(I)-stabilizing ligands in promoting protein labeling within the cytoplasm of E. coli, here we identify a highly efficient and biocompatible catalyst for intracellular modification of proteins by azide-alkyne cycloaddition. This reaction permits us to conjugate an environment-sensitive fluorophore site-specifically onto HdeA, an acid-stress chaperone that adopts pH-dependent conformational changes, in both the periplasm and cytoplasm of E. coli. The resulting protein-fluorophore hybrid pH indicators enable compartment-specific pH measurement to determine the pH gradient across the E. coli cytoplasmic membrane. This construct also allows the measurement of E. coli transmembrane potential, and the determination of the proton motive force across its inner membrane under normal and acid-stress conditions. 2014-09-19 /pmc/articles/PMC4174402/ /pubmed/25236616 http://dx.doi.org/10.1038/ncomms5981 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Yang, Maiyun
Jalloh, Abubakar S.
Wei, Wei
Zhao, Jing
Wu, Peng
Chen, Peng R.
Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli
title Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli
title_full Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli
title_fullStr Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli
title_full_unstemmed Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli
title_short Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli
title_sort biocompatible click chemistry enabled compartment-specific ph measurement inside e. coli
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4174402/
https://www.ncbi.nlm.nih.gov/pubmed/25236616
http://dx.doi.org/10.1038/ncomms5981
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