Single-Molecule Analysis of Glycopeptide Antibiotic-Target Interactions Using Nanopore-Based Biomimetic Probes

Area of Science:

• Biochemistry and Biophysics
• Microbiology and Infectious Diseases
• Nanotechnology and Single-Molecule Analysis

Background:

• Glycopeptide antibiotics are crucial for treating resistant bacterial infections.
• Emerging resistance, driven by target interaction changes, threatens their efficacy.
• Understanding these molecular interactions is key to developing new therapies.

Purpose of the Study:

• To develop a single-molecule method for studying glycopeptide-peptidoglycan interactions.
• To elucidate glycopeptide resistance mechanisms at the molecular level.
• To guide the design of next-generation glycopeptide antimicrobial agents.

Main Methods:

• Utilized a nanopore-based biomimetic probe strategy.
• Designed biomimetic peptidoglycan precursor peptides (BPGPPs).
• Employed the α-hemolysin (α-HL) nanopore for real-time single-molecule recordings of complex dissociation.

Main Results:

• Achieved real-time single-molecule recordings of glycopeptide-BPGPP complex dissociation.
• Binding affinity correlated with mean dwell time within the nanopore.
• Investigated mutations in BPGPPs to understand resistance mechanisms.

Conclusions:

• The nanopore method provides time- and space-resolved single-molecule insights into glycopeptide-peptidoglycan interactions.
• This approach aids in understanding natural resistance mechanisms.
• The findings support the development of novel antimicrobial agents against resistant bacteria.