High-throughput and facile assay of antimicrobial peptides using pH-controlled fluorescence resonance energy transfer
SCIE
SCOPUS
- Title
- High-throughput and facile assay of antimicrobial peptides using pH-controlled fluorescence resonance energy transfer
- Authors
- Kim, YS; Cha, HJ
- Date Issued
- 2006-10
- Publisher
- AMER SOC MICROBIOLOGY
- Abstract
- Amphipathic antimicrobial peptides can destroy bacteria cells by inducing membrane permeabilization, forming one strategy for innate defense by various organisms. However, although the antimicrobial peptides are considered a promising alternative for use against multidrug-resistant bacteria, large-scale screening of potential candidate antimicrobial peptides will require a simple, rapid assay for antimicrobial activity. Here, we describe a novel fluorescence resonance energy transfer (FRET)-based assay system for antimicrobial peptides which takes advantage of pH-related changes in FRET efficiency due to the instability of enhanced yellow fluorescent protein versus the stability of enhanced cyan fluorescent protein in a reduced-pH environment. We successfully showed that quantification of antimicrobial activity is possible through a difference of FRET efficiency between ECFP-EYFP fusion molecules released from disrupted Escherichia coli in an extracellular environment (pH 6) and those retained in an intracellular environment (pH similar to 7). Thus, we herein suggest a new simple, effective, and efficient pH-controlled FRET-based antimicrobial peptide screening method applicable to high-throughput screening of candidate peptide libraries.
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/9343
- DOI
- 10.1128/AAC.00455-06
- ISSN
- 0066-4804
- Article Type
- Article
- Citation
- ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, vol. 50, no. 10, page. 3330 - 3335, 2006-10
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