Structural characterization of m. tuberculosis IGP synthase (IGPS) variants using circular dichroism (CD) spectroscopy
Presentation Type
Abstract
Faculty Advisor
Nina Goodey
Access Type
Event
Start Date
25-4-2025 12:00 PM
End Date
25-4-2025 1:00 PM
Description
Tuberculosis is a deadly disease that has plagued the world for years and is caused by the pathogen Mycobacterium tuberculosis. Tuberculosis has grown resistant to various antibiotics over time, and poses a threat to the health of millions of people. It has been discovered that during the 4th step of the tryptophan biosynthetic pathway, the substrate CdRP is converted to IGP. This process is catalyzed by the enzyme Indole-3-Glycerol Phosphate Synthase (IGPS), and is essential for the growth and survival of the TB causing bacteria, which could be studied as a potential drug target. This study aims to characterize the structural properties of wildtype IGPS and its mutants (E57D and E219D) using circular dichroism (CD) spectroscopy, which will provide insight into how these mutations affect protein secondary structure and stability. The wildtype and mutant (E57D, E219D) IGPS proteins were expressed and purified using His-tag affinity chromatography. Structural changes between the wildtype and mutants will be analyzed using CD spectroscopy under different concentrations of glycerol. These results can potentially help explain why the E57D and E219D variants have higher catalytic activities in more viscous solvents. Future studies will investigate how these structural changes are involved in IGPS function and inhibitor binding, which may inform drug development efforts for tuberculosis treatment.
Structural characterization of m. tuberculosis IGP synthase (IGPS) variants using circular dichroism (CD) spectroscopy
Tuberculosis is a deadly disease that has plagued the world for years and is caused by the pathogen Mycobacterium tuberculosis. Tuberculosis has grown resistant to various antibiotics over time, and poses a threat to the health of millions of people. It has been discovered that during the 4th step of the tryptophan biosynthetic pathway, the substrate CdRP is converted to IGP. This process is catalyzed by the enzyme Indole-3-Glycerol Phosphate Synthase (IGPS), and is essential for the growth and survival of the TB causing bacteria, which could be studied as a potential drug target. This study aims to characterize the structural properties of wildtype IGPS and its mutants (E57D and E219D) using circular dichroism (CD) spectroscopy, which will provide insight into how these mutations affect protein secondary structure and stability. The wildtype and mutant (E57D, E219D) IGPS proteins were expressed and purified using His-tag affinity chromatography. Structural changes between the wildtype and mutants will be analyzed using CD spectroscopy under different concentrations of glycerol. These results can potentially help explain why the E57D and E219D variants have higher catalytic activities in more viscous solvents. Future studies will investigate how these structural changes are involved in IGPS function and inhibitor binding, which may inform drug development efforts for tuberculosis treatment.
Comments
Poster presentation at the 2025 Student Research Symposium.