Journal article
Adenylate Kinase Release as a High-Throughput-Screening-Compatible Reporter of Bacterial Lysis for Identification of Antibacterial Agents
Antimicrobial agents and chemotherapy, Vol.57(1), pp.26-36
01/2013
DOI: 10.1128/AAC.01640-12
PMCID: PMC3535927
PMID: 23027196
Abstract
Adenylate kinase (AK) is a ubiquitous intracellular enzyme that is released into the extracellular space upon cell lysis. We have shown that AK release serves as a useful reporter of bactericidal agent activity and can be exploited for antimicrobial screening purposes. The AK assay exhibits improved sensitivity over that of growth-based assays and can detect agents that are active against bacteria in clinically relevant growth states that are difficult to screen using conventional approaches, such as small colony variants (SCV) and bacteria within established biofilms. The usefulness of the AK assay was validated by screening a library of off-patent drugs for agents that exhibit antimicrobial properties toward a variety of bacterial species, including
Escherichia coli
and all members of the “ESKAPE” pathogens (E
nterococcus faecium
, S
taphylococcus aureus
, K
lebsiella pneumoniae
, A
cinetobacter baumannii
, P
seudomonas aeruginosa
, and E
nterobacter
species). The assay detected antibiotics within the library that were expected to be active against the organism screened. Moreover, 38 drugs with no previously reported antibacterial activity elicited AK release. Four of these were acquired, and all were verified to exhibit antimicrobial activity by standard susceptibility testing. Two of these molecules were further characterized. The antihistamine, terfenadine, was active against
S. aureus
planktonic, SCV population, and biofilm-associated cells. Tamoxifen, an estrogen receptor antagonist, was active toward
E. faecium
in vitro
and also reduced
E. faecium
pathogenesis in a
Galleria mellonella
infection model. Our data demonstrate that the AK assay provides an attractive screening approach for identifying new antimicrobial agents. Further, terfenadine and tamoxifen may represent novel antimicrobial drug development scaffolds.
Details
- Title: Subtitle
- Adenylate Kinase Release as a High-Throughput-Screening-Compatible Reporter of Bacterial Lysis for Identification of Antibacterial Agents
- Creators
- Anna C Jacobs - Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USALouis DiDone - Department of Pediatrics, University of Rochester School of Medicine and Dentistry, Rochester, New York, USAJennielle Jobson - Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USAMadeline K Sofia - Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USADamian Krysan - Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USAPaul M Dunman - Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USA
- Resource Type
- Journal article
- Publication Details
- Antimicrobial agents and chemotherapy, Vol.57(1), pp.26-36
- Publisher
- American Society for Microbiology
- DOI
- 10.1128/AAC.01640-12
- PMID
- 23027196
- PMCID
- PMC3535927
- ISSN
- 0066-4804
- eISSN
- 1098-6596
- Language
- English
- Date published
- 01/2013
- Academic Unit
- Microbiology and Immunology; Stead Family Department of Pediatrics; Infectious Disease (Pediatrics)
- Record Identifier
- 9984093212102771
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