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Screening for FtsZ Dimerization Inhibitors Using Fluorescence Cross-Correlation Spectroscopy and Surface Resonance

Shintaro Mikuni¹, Kota Kodama², Akira Sasaki³

¹Laboratory of Molecular Cell Dynamics, Faculty of Advanced Life Science, Hokkaido University, Sapporo, Japan.

|July 9, 2015

Summary

Researchers identified novel chemical compounds targeting FtsZ, a key bacterial cell division protein. This discovery offers a new strategy for developing antibiotics against bacteria, including drug-resistant strains like MRSA.

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Area of Science:

Microbiology
Biochemistry
Drug Discovery

Background:

FtsZ is an essential bacterial cell division protein and a promising target for novel antibiotics.
GTP-dependent polymerization of FtsZ is crucial for bacterial cell division.

Purpose of the Study:

To develop a high-throughput screening method for identifying FtsZ inhibitors.
To discover seed chemical structures with antibacterial activity against Staphylococcus aureus.

Main Methods:

Utilized fluorescence cross-correlation spectroscopy (FCCS) and surface plasmon resonance (SPR) for quantitative screening.
Developed a novel FCCS application using fragmented, fluorescently labeled FtsZ (Staphylococcus aureus FtsZ fused with GFP and mCherry).
Performed in silico screening and structural similarity searches to select chemical candidates.

Main Results:

Identified 28 and 71 chemical candidates through two rounds of FCCS screening.
Determined dissociation constants of top inhibitory chemicals using SPR.
Confirmed antibacterial activity of screened compounds against Staphylococcus aureus, including methicillin-resistant Staphylococcus aureus (MRSA).

Conclusions:

The developed FCCS-based method is effective for screening polymerization-prone proteins like FtsZ.
Identified potent FtsZ inhibitors with confirmed antibacterial activity.
These findings provide a foundation for developing new antibiotics targeting bacterial cell division.