Measuring Activity of ITPase P32T Variant through High-Performance Liquid Chromatography
Faculty Mentor
Nicholas Burgis
Presentation Type
Poster
Start Date
4-14-2026 9:00 AM
End Date
4-14-2026 11:00 AM
Location
PUB NCR
Primary Discipline of Presentation
Chemistry and Biochemistry
Abstract
ITPase, a pyrophosphohydrolase, is an enzyme that is responsible for the conversion of ITP (inosine triphosphate) into IMP (inosine monophosphate). ITP is a high-energy nucleotide, similar to ATP. One clinically relevant variation of this enzyme, called the P32T variant, is expressed in up to 10% of the population and causes complications to a class of drugs called thiopurines, used in some chemotherapy and immunosuppressant treatments. This summer, the P32T variant will potentially be included in a drug screening test, and in fall 2026 stabilization testing with small molecules will be conducted. The primary purpose of this experiment was to confirm the activity of ITPase with the P32T variation, under different conditions and using more sophisticated methods. In this experiment, high-performance liquid chromatography (HPLC) assays were used to determine the activity of the enzyme. Michaelis Menton kinetics, determined by using different concentrations of the substrate (ITP) will indicate how the enzyme handles the conversion of ITP into IMP. Data analysis of P32T and wildtype variants will be discussed.
Recommended Citation
Chamberlin, Alice, "Measuring Activity of ITPase P32T Variant through High-Performance Liquid Chromatography" (2026). 2026 Symposium. 14.
https://dc.ewu.edu/srcw_2026/ps_2026/p1_2026/14
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Measuring Activity of ITPase P32T Variant through High-Performance Liquid Chromatography
PUB NCR
ITPase, a pyrophosphohydrolase, is an enzyme that is responsible for the conversion of ITP (inosine triphosphate) into IMP (inosine monophosphate). ITP is a high-energy nucleotide, similar to ATP. One clinically relevant variation of this enzyme, called the P32T variant, is expressed in up to 10% of the population and causes complications to a class of drugs called thiopurines, used in some chemotherapy and immunosuppressant treatments. This summer, the P32T variant will potentially be included in a drug screening test, and in fall 2026 stabilization testing with small molecules will be conducted. The primary purpose of this experiment was to confirm the activity of ITPase with the P32T variation, under different conditions and using more sophisticated methods. In this experiment, high-performance liquid chromatography (HPLC) assays were used to determine the activity of the enzyme. Michaelis Menton kinetics, determined by using different concentrations of the substrate (ITP) will indicate how the enzyme handles the conversion of ITP into IMP. Data analysis of P32T and wildtype variants will be discussed.
