Kinetic Analysis of Yersinia pestis DNA Adenine Methyltransferase Activity
Author Information
Author(s): Wood Robert J., Maynard-Smith Michael D., Robinson Victoria L., Oyston Petra C.F., Titball Rick W., Roach Peter L.
Primary Institution: School of Chemistry, University of Southampton
Hypothesis
Selective Dam inhibitors might function as broad spectrum antibiotics.
Conclusion
The developed assay is suitable for high throughput screening for inhibitors of DNA adenine methyltransferases.
Supporting Evidence
- The assay has been validated for high throughput screening, giving a Z-factor of 0.71±0.07.
- Manipulating the levels of Dam in bacteria has been shown to reduce bacterial virulence.
- The developed assay allows for real-time monitoring of Dam methylation.
- Y. pestis Dam was observed to be sensitive towards inactivation.
Takeaway
Scientists created a new test to see how a specific enzyme works, which could help find new antibiotics.
Methodology
The study involved expressing and purifying Yersinia pestis Dam and developing a continuous fluorescence-based assay for its activity.
Limitations
The assay's initial rate determination was complicated by the enzyme's instability without substrates.
Statistical Information
P-Value
6.93±2.01 µM
Confidence Interval
0.71±0.07
Statistical Significance
p<0.05
Digital Object Identifier (DOI)
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