Abstract: —2′-Deoxyuridine 5′-triphosphate nucleotide hhydrolase (Dut) hydrolyzes dUTP to dUMP and pyrophosphate to prevent erroneous incorporation of dUMP from the dUTP metabolic pool into DNA. Dut is considered as a promising pharmacological target for antimetabolite therapy. Enzymatically active Dut is a trimer that binds the substrate at the interface between the subunits. High-speed nanoscale differential scanning fluorimetry (nanoDSF) was used to study how various physicochemical factors affect the stability of the Escherichia coli Dut trimer. Unlike with monomeric proteins, thermal unfolding of Dut occurred in two steps, the first one corresponding to dissociation of the trimer into monomeric subunits. Hydrophobic interactions and hydrogen bonds at the interfaces between the subunits were found to contribute most to trimer stabilization. The binding of nucleotide ligands partly stabilized the Dut trimer. In general, nanoDSF is a convenient assay for screening low-molecular-weight compounds for their ability to destabilize the active Dut trimer

Cite: Yudkina A.V. , Kovalenko E.A. , Endutkin A.V. , Panferova E.P. , Kirilenko A.A. , Kokhanenko A.A. , Zharkov D.O.
Factors Affecting the Stability of the Trimer of 2'-Deoxyuridine 5'-Triphosphate Nucleotide Hydrolase from Escherichia coli
Molecular Biology. 2023. V.57. N2. P.312-319. DOI: 10.1134/s002689332302022x WOS Scopus OpenAlex

Original: Юдкина А.В. , Коваленко Е.А. , Ендуткин А.В. , Панферова Е.П. , Кириленко А.А. , Коханенко А.А. , Жарков Д.О.
Факторы, влияющие на стабильность тримерной формы 2′-дезоксиуридин-5′-трифосфатнуклеотидгидролазы Escherichia coli
Молекулярная биология. 2023. Т.57. №2. С.330-339. DOI: 10.31857/S0026898423020246 РИНЦ OpenAlex

Dates:

Published print:

Aug 16, 2022

Identifiers:

Web of science:	WOS:000984409600016
Scopus:	2-s2.0-85156136183
OpenAlex:	W4367056758

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DB	Citing
OpenAlex	1
Scopus	1

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