Abstract
Inorganic polyphosphate (polyP) and its metabolic enzymes are important in several cellular processes related with virulence and antibiotic susceptibility. Accordingly, bacterial polyP synthesis has been proposed as a good target for designing novel antivirulence molecules as alternative to conventional antibiotics. In most pathogenic bacteria, polyphosphate kinase 1 (PPK1), in charge of polyP synthesis from ATP, is widely conserved. Current colorimetric and radioactive polyP synthesis enzymatic assays are not suitable for high-throughput screening of PPK1 inhibitors. Given the ability of polyP to modify the excitation-emission spectra of DAPI (4ʹ-6-diamidino-2-phenylindole), a fluorescence assay was previously developed by using a purified recombinant PPK1 enzyme from Escherichia coli. In this work we have developed a suitable methodology for high-throughput measurement of E. coli PPK1 activity. This platform can be used for the screening putative antimicrobial molecules for related enteropathogenic bacteria.
Original language | English |
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Pages (from-to) | 2237-2242 |
Number of pages | 6 |
Journal | Infection and Drug Resistance |
Volume | 12 |
DOIs | |
State | Published - 2019 |
Bibliographical note
Publisher Copyright:© 2019 Campos et al.
ASJC Scopus subject areas
- Pharmacology
- Infectious Diseases
- Pharmacology (medical)