Event Title

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.

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Apr 22nd, 10:25 AM Apr 22nd, 10:40 AM

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.