In vivo Testing of a Lethal ITPA Mutant
Faculty Mentor
Nicholas Burgis
Presentation Type
Poster
Start Date
May 2025
End Date
May 2025
Location
PUB NCR
Primary Discipline of Presentation
Chemistry and Biochemistry
Abstract
The inosine triphosphate pyrophosphatase (ITPA) protein is responsible for removing noncanonical purines, such as inosine triphosphate (ITP), from the intracellular nucleoside triphosphate pools. This prevents ITP from accumulating in the cell and disrupting cellular processes. A point mutation in ITPA at position 178 (R178C) results in an enzyme with severely reduced catalytic activity and stability, which causes a fatal infantile encephalopathy. This mutation was cloned into a plasmid that was transformed into Escherichia coli, creating an in vivo complementation model. To characterize the mutant, two complementation assays were performed to determine temperature and N-6 hydroxyaminopurine (HAP) sensitivity. This in vivo model of R178C can be used to test if druglike molecules are able to restore the functionality of the enzyme.
Recommended Citation
Burgis NE, April C, VanWormer K. Arginine-178 is an essential residue for ITPA function. Arch Biochem Biophys. 2023 Aug;744:109700. doi: 10.1016/j.abb.2023.109700. Epub 2023 Jul 26. PMID: 37506994; PMCID: PMC10530447. Burgis NE. A disease spectrum for ITPA variation: advances in biochemical and clinical research. J Biomed Sci. 2016 Oct 22;23(1):73. doi: 10.1186/s12929-016-0291-y. PMID: 27770805; PMCID: PMC5075207.
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
In vivo Testing of a Lethal ITPA Mutant
PUB NCR
The inosine triphosphate pyrophosphatase (ITPA) protein is responsible for removing noncanonical purines, such as inosine triphosphate (ITP), from the intracellular nucleoside triphosphate pools. This prevents ITP from accumulating in the cell and disrupting cellular processes. A point mutation in ITPA at position 178 (R178C) results in an enzyme with severely reduced catalytic activity and stability, which causes a fatal infantile encephalopathy. This mutation was cloned into a plasmid that was transformed into Escherichia coli, creating an in vivo complementation model. To characterize the mutant, two complementation assays were performed to determine temperature and N-6 hydroxyaminopurine (HAP) sensitivity. This in vivo model of R178C can be used to test if druglike molecules are able to restore the functionality of the enzyme.
