A metal‐dependent conformational change provides a structural basis for the inhibition of CTP synthase by gemcitabine‐5′‐triphosphate

McLeod, Matthew; Tran, Norman; McCluskey, Gregory D.; Gillis, Tom D; Bearne, Stephen L.; Holyoak, Todd

doi:10.1002/pro.4648

Cited by 5 publications

(1 citation statement)

References 58 publications

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“…Recently, McLeod et al unveiled the inhibitory effects of gemcitabine-5′triphosphate, a therapeutic agent against solid tumors, on ecCTPS 30 . It exhibited a strong binding affinity to CTPS, approximately 80-fold greater than CTP, providing novel insights into inhibitor development 31 . Utilizing cryo-electron microscopy (cryo-EM), Lynch et al revealed the mechanisms of selective inhibition of human CTPS and laid a foundation for the design of immunosuppressive therapies 17 .…”

Section: Introductionmentioning

confidence: 99%

Filamentation and inhibition of prokaryotic CTP synthase with ligands

Guo,

Wang,

Liu

2024

mLife

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Cytidine triphosphate synthase (CTPS) plays a pivotal role in the de novo synthesis of cytidine triphosphate (CTP), a fundamental building block for RNA and DNA that is essential for life. CTPS is capable of directly binding to all four nucleotide triphosphates: adenine triphosphate, uridine triphosphate, CTP, and guanidine triphosphate. Furthermore, CTPS can form cytoophidia in vivo and metabolic filaments in vitro, undergoing regulation at multiple levels. CTPS is considered a potential therapeutic target for combating invasions or infections by viral or prokaryotic pathogens. Utilizing cryo‐electron microscopy, we determined the structure of Escherichia coli CTPS (ecCTPS) filament in complex with CTP, nicotinamide adenine dinucleotide (NADH), and the covalent inhibitor 6‐diazo‐5‐oxo‐ l‐norleucine (DON), achieving a resolution of 2.9 Å. We constructed a phylogenetic tree based on differences in filament‐forming interfaces and designed a variant to validate our hypothesis, providing an evolutionary perspective on CTPS filament formation. Our computational analysis revealed a solvent‐accessible ammonia tunnel upon DON binding. Through comparative structural analysis, we discern a distinct mode of CTP binding of ecCTPS that differs from eukaryotic counterparts. Combining biochemical assays and structural analysis, we determined and validated the synergistic inhibitory effects of CTP with NADH or adenine on CTPS. Our results expand our comprehension of the diverse regulatory aspects of CTPS and lay a foundation for the design of specific inhibitors targeting prokaryotic CTPS.

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