Research output: Contribution to journal › Article › peer-review
Factors Affecting the Stability of the Trimer of 2'-Deoxyuridine 5'-Triphosphate Nucleotide Hydrolase from Escherichia coli. / Yudkina, A. V.; Kovalenko, E. A.; Endutkin, A. V. et al.
In: Molecular Biology, Vol. 57, No. 2, 04.2023, p. 312-319.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Factors Affecting the Stability of the Trimer of 2'-Deoxyuridine 5'-Triphosphate Nucleotide Hydrolase from Escherichia coli
AU - Yudkina, A. V.
AU - Kovalenko, E. A.
AU - Endutkin, A. V.
AU - Panferova, E. P.
AU - Kirilenko, A. A.
AU - Kokhanenko, A. A.
AU - Zharkov, D. O.
N1 - This work was supported by the Strategic academic leadership program of Tomsk State University (Priority-2030). Structural analysis was supported by the Ministry of Science and Higher Education of the Russian Federation (agreement no. 075-15-2022-263). Публикация для корректировки.
PY - 2023/4
Y1 - 2023/4
N2 - 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.
AB - 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.
KW - dUTPase
KW - differential scanning fluorimetry
KW - oligomerization
KW - protein–protein interactions
UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85156136183&origin=inward&txGid=b7a56ad9236324d4d47eab88a55ea93a
UR - https://www.mendeley.com/catalogue/510dc72a-4746-309e-bf22-1434d55eb199/
U2 - 10.1134/S002689332302022X
DO - 10.1134/S002689332302022X
M3 - Article
VL - 57
SP - 312
EP - 319
JO - Molecular Biology
JF - Molecular Biology
SN - 0026-8933
IS - 2
ER -
ID: 59649130