Studies on the catalytic mechanism of five DNA glycosylases: Probing for enzyme-DNA imino intermediates

Bin Sun, Katherine A. Latham, M. L. Dodson, Robert (Stephen) Lloyd

Research output: Contribution to journalArticle

127 Citations (Scopus)

Abstract

DNA glycosylases catalyze scission of the N-glycosylic bond linking a damaged base to the DNA sugar phosphate backbone. Some of these enzymes carry out a concomitant abasic (apyrimidinic/apurinic (AP)) lyase reaction at a rate approximately equal to that of the glycosylase step. As a generalization of the mechanism described for T4 endonuclease V, a repair glycosylase/AP lyase that is specific for ultraviolet light-induced cis-syn pyrimidine dimers, a hypothesis concerning the mechanism of these repair glycosylases has been proposed. This hypothesis describes the initial action of all DNA glycosylases as a nucleophilic attack at the sugar C-1′ of the damaged base nucleoside, resulting in scission of the N-glycosylic bond. It is proposed that the enzymes that are only glycosylases differ in the chemical nature of the attacking nucleophile from the glycosylase/AP lyases. Those DNA glycosylases, which carry out the AP lyase reaction at a rate approximately equal to the glycosylase step, are proposed to use an amino group as the nucleophile, resulting in an imino enzyme-DNA intermediate. The simple glycosylases, lacking the concomitant AP lyase activity, are proposed to use some nucleophile from the medium, e.g. an activated water molecule. This paper reports experimental tests of this hypothesis using five representative enzymes, and these data are consistent with this hypothesis.

Original languageEnglish (US)
Pages (from-to)19501-19508
Number of pages8
JournalJournal of Biological Chemistry
Volume270
Issue number33
StatePublished - Aug 18 1995
Externally publishedYes

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DNA Glycosylases
Lyases
Nucleophiles
DNA
Enzymes
Repair
Sugar Phosphates
Pyrimidine Dimers
Ultraviolet Rays
Nucleosides
Sugars
Molecules
Water

ASJC Scopus subject areas

  • Biochemistry

Cite this

Studies on the catalytic mechanism of five DNA glycosylases : Probing for enzyme-DNA imino intermediates. / Sun, Bin; Latham, Katherine A.; Dodson, M. L.; Lloyd, Robert (Stephen).

In: Journal of Biological Chemistry, Vol. 270, No. 33, 18.08.1995, p. 19501-19508.

Research output: Contribution to journalArticle

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