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

Maiyun Yang, Abubakar S. Jalloh, Wei Wei, Jing Zhao, Peng Wu, Peng R. Chen

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Bioorthogonal reactions, especially the Cu(I)-catalysed 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 labelling within the cytoplasm of Escherichia coli, 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.

Original languageEnglish (US)
Article number4981
JournalNature Communications
Volume5
DOIs
StatePublished - 2014
Externally publishedYes

Fingerprint

Click Chemistry
compartments
Escherichia coli
chemistry
Proton-Motive Force
Alkynes
Azides
Cycloaddition
Fluorophores
cytoplasm
Cycloaddition Reaction
cycloaddition
alkynes
proteins
Cytoplasm
membranes
Membranes
Periplasm
acids
Proteins

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry(all)
  • Physics and Astronomy(all)

Cite this

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.

In: Nature Communications, Vol. 5, 4981, 2014.

Research output: Contribution to journalArticle

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. In: Nature Communications. 2014 ; Vol. 5.
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