Investigation of Bacterial Carbonic Anhydrase Stability
Session Number
Project ID: BIO 01
Advisor(s)
Dr. James R. Horn; Northern Illinois University
Dakota Grote; Northern Illinois University
Discipline
Biology
Start Date
22-4-2020 10:25 AM
End Date
22-4-2020 10:40 AM
Abstract
The purpose of this study was to investigate the stability of Bartonella henselae carbonic anhydrase in an effort to design screening methods for potential inhibitors. Although several carbonic anhydrase inhibitors, such as acetazolamide and ethoxzolamide, are marketed drugs, new carbonic anhydrase inhibitors may be used to potentially combat bacterial infections. In this study, the thermal stability of B. henselae carbonic anhydrase was evaluated using a fluorescence thermal shift (FTS) assay to determine the enzyme’s melting temperature (TM).
Data from the initial buffer screen showed that the Tm of B. henselae carbonic anhydrase was around 80°C under a variety of conditions. Due to the high Tm value, which can complicate FTS assays evaluating ligand binding, methods were explored to decrease the stability of carbonic anhydrase. Investigation of known inhibitors, acetazolamide and ethoxzolamide, was conducted to investigate their stabilization of carbonic anhydrase. The results of this study will help guide the development of an effective screening method for potential carbonic anhydrase inhibitors.
Investigation of Bacterial Carbonic Anhydrase Stability
The purpose of this study was to investigate the stability of Bartonella henselae carbonic anhydrase in an effort to design screening methods for potential inhibitors. Although several carbonic anhydrase inhibitors, such as acetazolamide and ethoxzolamide, are marketed drugs, new carbonic anhydrase inhibitors may be used to potentially combat bacterial infections. In this study, the thermal stability of B. henselae carbonic anhydrase was evaluated using a fluorescence thermal shift (FTS) assay to determine the enzyme’s melting temperature (TM).
Data from the initial buffer screen showed that the Tm of B. henselae carbonic anhydrase was around 80°C under a variety of conditions. Due to the high Tm value, which can complicate FTS assays evaluating ligand binding, methods were explored to decrease the stability of carbonic anhydrase. Investigation of known inhibitors, acetazolamide and ethoxzolamide, was conducted to investigate their stabilization of carbonic anhydrase. The results of this study will help guide the development of an effective screening method for potential carbonic anhydrase inhibitors.