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The Role of Active-Site Plasticity in Damaged-Nucleotide Recognition by Human Apurinic/Apyrimidinic Endonuclease APE1. / Bulygin, Anatoly A.; Kuznetsova, Alexandra A.; Vorobjev, Yuri N. и др.
в: Molecules, Том 25, № 17, 3940, 01.09.2020.Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
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TY - JOUR
T1 - The Role of Active-Site Plasticity in Damaged-Nucleotide Recognition by Human Apurinic/Apyrimidinic Endonuclease APE1
AU - Bulygin, Anatoly A.
AU - Kuznetsova, Alexandra A.
AU - Vorobjev, Yuri N.
AU - Fedorova, Olga S.
AU - Kuznetsov, Nikita A.
N1 - Publisher Copyright: © 2020 by the authors. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/9/1
Y1 - 2020/9/1
N2 - Human apurinic/apyrimidinic (AP) endonuclease APE1 hydrolyzes phosphodiester bonds on the 50 side of an AP-site, and some damaged nucleotides such as 1,N6-ethenoadenosine (εA), α-adenosine (αA), and 5,6-dihydrouridine (DHU). To investigate the mechanism behind the broad substrate specificity of APE1, we analyzed pre-steady-state kinetics of conformational changes in DNA and the enzyme during DNA binding and damage recognition. Molecular dynamics simulations of APE1 complexes with one of damaged DNA duplexes containing εA, αA, DHU, or an F-site (a stable analog of an AP-site) revealed the involvement of residues Asn229, Thr233, and Glu236 in the mechanism of DNA lesion recognition. The results suggested that processing of an AP-site proceeds faster in comparison with nucleotide incision repair substrates because eversion of a small abasic site and its insertion into the active site do not include any unfavorable interactions, whereas the insertion of any target nucleotide containing a damaged base into the APE1 active site is sterically hindered. Destabilization of the α-helix containing Thr233 and Glu236 via a loss of the interaction between these residues increased the plasticity of the damaged-nucleotide binding pocket and the ability to accommodate structurally different damaged nucleotides. Nonetheless, the optimal location of εA or αA in the binding pocket does not correspond to the optimal conformation of catalytic amino acid residues, thereby significantly decreasing the cleavage efficacy for these substrates.
AB - Human apurinic/apyrimidinic (AP) endonuclease APE1 hydrolyzes phosphodiester bonds on the 50 side of an AP-site, and some damaged nucleotides such as 1,N6-ethenoadenosine (εA), α-adenosine (αA), and 5,6-dihydrouridine (DHU). To investigate the mechanism behind the broad substrate specificity of APE1, we analyzed pre-steady-state kinetics of conformational changes in DNA and the enzyme during DNA binding and damage recognition. Molecular dynamics simulations of APE1 complexes with one of damaged DNA duplexes containing εA, αA, DHU, or an F-site (a stable analog of an AP-site) revealed the involvement of residues Asn229, Thr233, and Glu236 in the mechanism of DNA lesion recognition. The results suggested that processing of an AP-site proceeds faster in comparison with nucleotide incision repair substrates because eversion of a small abasic site and its insertion into the active site do not include any unfavorable interactions, whereas the insertion of any target nucleotide containing a damaged base into the APE1 active site is sterically hindered. Destabilization of the α-helix containing Thr233 and Glu236 via a loss of the interaction between these residues increased the plasticity of the damaged-nucleotide binding pocket and the ability to accommodate structurally different damaged nucleotides. Nonetheless, the optimal location of εA or αA in the binding pocket does not correspond to the optimal conformation of catalytic amino acid residues, thereby significantly decreasing the cleavage efficacy for these substrates.
KW - 5,6-dihydrouridine
KW - Active site plasticity
KW - AP endonuclease
KW - Apurinic/apyrimidinic site
KW - Base excision repair
KW - Conformational dynamics
KW - apurinic
KW - PROTEIN
KW - base excision repair
KW - 6-dihydrouridine
KW - INCISION ACTIVITY
KW - GLYCOSYLASES
KW - active site plasticity
KW - apyrimidinic site
KW - conformational dynamics
KW - 5
KW - DIVALENT METAL-IONS
KW - KINETIC MECHANISM
KW - DYNAMICS
KW - CHEMISTRY
KW - BASE-EXCISION
KW - DNA-REPAIR
KW - BINDING
UR - http://www.scopus.com/inward/record.url?scp=85090182163&partnerID=8YFLogxK
U2 - 10.3390/molecules25173940
DO - 10.3390/molecules25173940
M3 - Article
C2 - 32872297
AN - SCOPUS:85090182163
VL - 25
JO - Molecules
JF - Molecules
SN - 1420-3049
IS - 17
M1 - 3940
ER -
ID: 25290340