Kinetic Characterization and Inhibition of Trypanosoma cruzi Hypoxanthine–Guanine Phosphoribosyltransferases

• Published in: Biochemistry (Easton) Vol. 61; no. 19; pp. 2088 - 2105
• Main Authors: Glockzin, Kayla, Kostomiris, Demetrios, Minnow, Yacoba V. T., Suthagar, Kajitha, Clinch, Keith, Gai, Sinan, Buckler, Joshua N., Schramm, Vern L., Tyler, Peter C., Meek, Thomas D., Katzfuss, Ardala
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 04.10.2022
• Subjects: Antiparasitic Agents; Chagas Disease; Guanine - metabolism; Guanosine Monophosphate; Humans; Hypoxanthine Phosphoribosyltransferase - chemistry; Hypoxanthine Phosphoribosyltransferase - genetics; Hypoxanthine Phosphoribosyltransferase - metabolism; Inosine Monophosphate; Isoenzymes; Purines - metabolism; Purines - pharmacology; Trypanosoma cruzi
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/acs.biochem.2c00312

Chagas disease, caused by the parasitic protozoan Trypanosoma cruzi, affects over 8 million people worldwide. Current antiparasitic treatments for Chagas disease are ineffective in treating advanced, chronic stages of the disease, and are noted for their toxicity. Like most parasitic protozoa, T. cruzi is unable to synthesize purines de novo, and relies on the salvage of preformed purines from the host. Hypoxanthine–guanine phosphoribosyltransferases (HGPRTs) are enzymes that are critical for the salvage of preformed purines, catalyzing the formation of inosine monophosphate (IMP) and guanosine monophosphate (GMP) from the nucleobases hypoxanthine and guanine, respectively. Due to the central role of HGPRTs in purine salvage, these enzymes are promising targets for the development of new treatment methods for Chagas disease. In this study, we characterized two gene products in the T. cruzi CL Brener strain that encodes enzymes with functionally identical HGPRT activities in vitro: TcA (TcCLB.509693.70) and TcC (TcCLB.506457.30). The TcC isozyme was kinetically characterized to reveal mechanistic details on catalysis, including identification of the rate-limiting step(s) of catalysis. Furthermore, we identified and characterized inhibitors of T. cruzi HGPRTs originally developed as transition-state analogue inhibitors (TSAIs) of Plasmodium falciparum hypoxanthine–guanine–xanthine phosphoribosyltransferase (PfHGXPRT), where the most potent compound bound to T. cruzi HGPRT with low nanomolar affinity. Our results validated the repurposing of TSAIs to serve as selective inhibitors for orthologous molecular targets, where primary and secondary structures as well as putatively common chemical mechanisms are conserved.