Microspectrofluorimetry to Dissect the Permeation of Ceftazidime in Gram-negative Bacteria

Overview

Journal Sci Rep

Specialty Science

Date 2017 Apr 22

PMID 28428543

Citations 9

Authors

Anas Allam

Laure Maigre

Julia Vergalli

Estelle Dumont

Bertrand Cinquin

Rodolphe Alves de Sousa

Jelena Pajovic

Elizabeth Pinet

Nikaia Smith

Jean-Philippe Herbeuval

Matthieu Refregiers

Isabelle Artaud

Jean-Marie Pages

Affiliations

Soon will be listed here.

Abstract

A main challenge in chemotherapy is to determine the in cellulo parameters modulating the drug concentration required for therapeutic action. It is absolutely urgent to understand membrane permeation and intracellular concentration of antibiotics in clinical isolates: passing the membrane barrier to reach the threshold concentration inside the bacterial periplasm or cytoplasm is the pivotal step of antibacterial activity. Ceftazidime (CAZ) is a key molecule of the combination therapy for treating resistant bacteria. We designed and synthesized different fluorescent CAZ derivatives (CAZ*, CAZ**) to dissect the early step of translocation-accumulation across bacterial membrane. Their activities were determined on E. coli strains and on selected clinical isolates overexpressing ß-lactamases. The accumulation of CAZ* and CAZ** were determined by microspectrofluorimetry and epifluorimetry. The derivatives were properly translocated to the periplasmic space when we permeabilize the outer membrane barrier. The periplasmic location of CAZ** was related to a significant antibacterial activity and with the outer membrane permeability. This study demonstrated the correlation between periplasmic accumulation and antibiotic activity. We also validated the method for approaching ß-lactam permeation relative to membrane permeability and paved the way for an original matrix for determining "Structure Intracellular Accumulation Activity Relationship" for the development of new therapeutic candidates.

Citing Articles

Cytoplasmic Accumulation and Permeability of Antibiotics in Gram Positive and Gram Negative Bacteria Visualized in Real-Time via a Fluorogenic Tagging Strategy.

Squire S, Sebghati S, Hammond M ACS Chem Biol. 2023; 19(1):3-8.

PMID: 38096425 PMC: 10805102. DOI: 10.1021/acschembio.3c00510.

Cephalosporin translocation across enterobacterial OmpF and OmpC channels, a filter across the outer membrane.

Masi M, Vergalli J, Ghai I, Barba-Bon A, Schembri T, Nau W Commun Biol. 2022; 5(1):1059.

PMID: 36198902 PMC: 9534850. DOI: 10.1038/s42003-022-04035-y.

Live-Cell Profiling of Penicillin-Binding Protein Inhibitors in MG1655.

Shirley J, Nauta K, Carlson E ACS Infect Dis. 2022; 8(7):1241-1252.

PMID: 35763562 PMC: 10040144. DOI: 10.1021/acsinfecdis.2c00004.

Porins and small-molecule translocation across the outer membrane of Gram-negative bacteria.

Vergalli J, Bodrenko I, Masi M, Moynie L, Acosta-Gutierrez S, Naismith J Nat Rev Microbiol. 2019; 18(3):164-176.

PMID: 31792365 DOI: 10.1038/s41579-019-0294-2.

Fluoroquinolone-derived fluorescent probes for studies of bacterial penetration and efflux.

Stone M, Masi M, Phetsang W, Pages J, Cooper M, Blaskovich M Medchemcomm. 2019; 10(6):901-906.

PMID: 31303987 PMC: 6596217. DOI: 10.1039/c9md00124g.

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