Activity-based ATP analog probes for bacterial histidine kinases

Area of Science:

• Microbiology
• Biochemistry
• Molecular Biology

Background:

• Histidine kinases (HKs) are sensor proteins central to prokaryotic two-component systems (TCSs).
• TCSs regulate cellular responses to environmental stimuli, with HKs playing roles in bacterial virulence and antibiotic resistance.
• Identifying stimuli that activate HKs remains a significant knowledge gap.

Purpose of the Study:

• To develop and validate methods for evaluating the efficacy of potential HK inhibitors.
• To establish techniques for determining the kinetic parameters of activity-based probes targeting HKs.
• To facilitate the elucidation of environmental stimuli that activate HKs.

Main Methods:

• Activity-based protein profiling (ABPP) for studying HK activity.
• Development of assays to measure the potency of putative HK inhibitors.
• Kinetic analysis of activity-based probes designed for HKs.

Main Results:

• Established methods for assessing the potency of novel HK inhibitors.
• Developed protocols for calculating kinetic parameters of HK-specific activity-based probes.
• Provided a framework for identifying environmental triggers of HK signaling.

Conclusions:

• HK inhibitors show promise as standalone antibiotics or antivirulence therapies.
• Activity-based protein profiling is a viable strategy to uncover stimuli activating HKs.
• This work advances the understanding and therapeutic targeting of bacterial TCSs.