Research output: Contribution to journal › Article › peer-review
Interaction features of adenine DNA glycosylase MutY from E. coli with DNA substrates. / Tyugashev, T. E.; Kuznetsova, A. A.; Kuznetsov, N. A. et al.
In: Russian Journal of Bioorganic Chemistry, Vol. 43, No. 1, 01.01.2017, p. 13-22.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Interaction features of adenine DNA glycosylase MutY from E. coli with DNA substrates
AU - Tyugashev, T. E.
AU - Kuznetsova, A. A.
AU - Kuznetsov, N. A.
AU - Fedorova, O. S.
PY - 2017/1/1
Y1 - 2017/1/1
N2 - Kinetic characteristics of specific recognition of damaged base by the DNA glycosylase MutY in model DNA substrates, containing oxoG/A-, G/A-, oxoG/C- and F/G pairs in the central position, were investigated. Conformational changes of the MutY enzyme during the recognition of the damaged base in DNA have been recorded by the change in the fluorescence intensity of tryptophan residues using the stopped-flow technique in real time. DNA duplexes containing a fluorescein residue were used for the registration of DNA conformational changes. Analysis of the kinetic curves allowed us to determine the values of rate constants for the kinetic stages of the interaction. It was shown that nonspecific contacts between the DNA-binding site of the enzyme and the DNA duplex are formed at the first stage of the interaction. It was found that the discrimination of Gua and oxoGua bases occurs at the second stage of the MutY interaction with the DNA duplex. The data obtained for the oxoG/C-substrate showed that the recognition of the base located opposite oxoGua also occurs at this stage.
AB - Kinetic characteristics of specific recognition of damaged base by the DNA glycosylase MutY in model DNA substrates, containing oxoG/A-, G/A-, oxoG/C- and F/G pairs in the central position, were investigated. Conformational changes of the MutY enzyme during the recognition of the damaged base in DNA have been recorded by the change in the fluorescence intensity of tryptophan residues using the stopped-flow technique in real time. DNA duplexes containing a fluorescein residue were used for the registration of DNA conformational changes. Analysis of the kinetic curves allowed us to determine the values of rate constants for the kinetic stages of the interaction. It was shown that nonspecific contacts between the DNA-binding site of the enzyme and the DNA duplex are formed at the first stage of the interaction. It was found that the discrimination of Gua and oxoGua bases occurs at the second stage of the MutY interaction with the DNA duplex. The data obtained for the oxoG/C-substrate showed that the recognition of the base located opposite oxoGua also occurs at this stage.
KW - adenine DNA glycosylase
KW - conformational changes
KW - enzyme specificity
KW - pre-steady state
KW - OXIDATION
KW - REPAIR ENZYME MUTY
KW - DOMAIN
KW - RECOGNITION
KW - MECHANISM
KW - SPECIFICITY
KW - BASE EXCISION
KW - 8-OXOGUANINE
KW - STRUCTURAL BASIS
KW - FPG PROTEIN
UR - http://www.scopus.com/inward/record.url?scp=85012907529&partnerID=8YFLogxK
U2 - 10.1134/S1068162017010101
DO - 10.1134/S1068162017010101
M3 - Article
AN - SCOPUS:85012907529
VL - 43
SP - 13
EP - 22
JO - Russian Journal of Bioorganic Chemistry
JF - Russian Journal of Bioorganic Chemistry
SN - 1068-1620
IS - 1
ER -
ID: 8681425