2. Academic Validation
  3. Biocompatible click chemistry enabled compartment-specific pH measurement inside E. coli

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

  • Nat Commun. 2014 Sep 19;5:4981. doi: 10.1038/ncomms5981.
Maiyun Yang 1,  Abubakar S Jalloh 2,  Wei Wei 3,  Jing Zhao 3,  Peng Wu 2,  Peng R Chen 4
Affiliations

Affiliations

  • 1 Synthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.
  • 2 Department of Biochemistry, Albert Einstein College of Medicine of Yeshiva University, 1300 Morris Park Avenue, Bronx, New York 10461, USA.
  • 3 State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Institute of Chemistry and BioMedical Sciences, Nanjing University, Nanjing 210093, China.
  • 4 1] Synthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China [2] Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China.
PMID: 25236616 DOI: 10.1038/ncomms5981
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.

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  • HY-128708
    吡咯赖氨酸类似物
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