8I9O

ecCTPS filament bound with CTP, NADH, DON

8I9O の概要

• エントリーDOI: 10.2210/pdb8i9o/pdb
• EMDBエントリー: 35278
• 分子名称: CTP synthase, ADENINE, 5-OXO-L-NORLEUCINE, ... (5 entities in total)
• 機能のキーワード: inhibitor, metabolic filament, ctp synthase, hydrolase
• 由来する生物種: Escherichia coli 'BL21-Gold(DE3)pLysS AG'
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 244422.18

構造登録者

Guo, C.J.,Liu, J.L. (登録日: 2023-02-07, 公開日: 2024-02-14, 最終更新日: 2024-07-24)

主引用文献

Guo, C.,Wang, Z.,Liu, J.L.
Filamentation and inhibition of prokaryotic CTP synthase with ligands.
Mlife, 3:240-250, 2024

PubMed Abstract: 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 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.

PubMed: 38948148
DOI: 10.1002/mlf2.12119

実験手法

ELECTRON MICROSCOPY (2.9 Å)