Fluorescence Polarization Binding Assay for Aspergillus fumigatus Virulence Factor UDP-Galactopyranose Mutase
Author Information
Author(s): Qi Jun, Oppenheimer Michelle, Sobrado Pablo
Primary Institution: Virginia Tech
Hypothesis
Inhibitors of UDP-galactopyranose mutase (UGM) can block the biosynthesis of galactofuranose, leading to potential treatments for Aspergillus fumigatus infections.
Conclusion
The study developed a fluorescence polarization assay that effectively identifies inhibitors of AfUGM, which could lead to new treatments for A. fumigatus-related diseases.
Supporting Evidence
- UDP-TAMRA analogs bind to AfUGM with high affinities, suggesting they are effective probes for the enzyme.
- The fluorescence polarization assay showed a Z' factor of 0.79 ± 0.02, indicating good assay quality.
- Compounds 7 and 8 were tested and showed varying degrees of inhibition on AfUGM activity.
- The study identified that UDP-Galp is a poor ligand for AfUGM with a Kd value of 495 ± 66 μM.
Takeaway
The researchers created a test to find new medicines that can stop a fungus from making a part it needs to cause sickness.
Methodology
The study involved synthesizing fluorescently labeled UDP derivatives and developing a fluorescence polarization binding assay to measure their binding affinities to AfUGM.
Limitations
The binding affinities of the fluorescent probes to AfUGM were lower than expected, which may limit their use in high-throughput screening.
Statistical Information
P-Value
9.0 ± 1.7 μM
Confidence Interval
2.6 ± 0.2 μM
Statistical Significance
p<0.05
Digital Object Identifier (DOI)
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