Spectroscopic analysis of the pyrimidine(6-4)pyrimidone photoproduct: Insights into the (6-4) photolyase reaction

Junpei Yamamoto, Yoshiyuki Tanaka, Shigenori Iwai

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

We synthesized a dinucleoside monophosphate of the 15N-labeled (6-4) photoproduct, which is one of the major UV-induced lesions in DNA, to investigate the (6-4) photolyase repair mechanism, and characterized its protonation state by measuring 15N NMR spectra as a function of pH. We expected that chemical-shift changes of the pyrimidone 15N3, due to protonation, would be observed at pH 3, as observed for the 15N-labeled 5-methylpyrimidin-2-one nucleoside. Interestingly, however, the changes were observed only in alkaline solutions. In UV absorption spectroscopy and HPLC analyses under acidic conditions, a change in the maximum absorption wavelength, due to the protonation-induced hydrolysis, was observed at and below pH 1, but not at pH 2, whereas the protonation of 5-methylpyrimidin-2-one occurred at pH values between 2 and 3. These results indicated that the pKa value for this N3 is remarkably lower than that of a normal pyrimidone ring, and strongly suggest that an intramolecular hydrogen bond is formed between the N3 of the 3′ base and the 5-OH of the 5′ base under physiological conditions. The results of this study have implications not only for the recognition and reaction mechanisms of (6-4) photolyase, but also for the chemical nature of the (6-4) photoproduct.

Original languageEnglish
Pages (from-to)161-166
Number of pages6
JournalOrganic and Biomolecular Chemistry
Volume7
Issue number1
DOIs
Publication statusPublished - 2009 Jan 1

ASJC Scopus subject areas

  • Biochemistry
  • Physical and Theoretical Chemistry
  • Organic Chemistry

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