Repair activities of 8-oxoguanine DNA glycosylase from Archaeoglobus fulgidus, a hyperthermophilic archaeon

• Published in: Mutation research Vol. 486; no. 2; pp. 99 - 111
• Main Authors: Hyung Chung, Ji, Suh, Moo-Jin, In Park, Young, Tainer, John A., Sun Han, Ye
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 12.07.2001; Elsevier
• Subjects: 8-Oxoguanine; 8-Oxoguanine DNA glycosylase; Amino Acid Sequence; Apurinic/apyrimidinic (AP) lyase; apurinic/apyrimidinic lyase; Archaeal Proteins - chemistry; Archaeal Proteins - metabolism; Archaeoglobus fulgidus; Archaeoglobus fulgidus - enzymology; Archaeoglobus fulgidus - genetics; Bacteriology; Biological and medical sciences; DNA glycosylase; DNA Repair; DNA-Formamidopyrimidine Glycosylase; Enzyme Stability; Escherichia coli; Escherichia coli Proteins; Fundamental and applied biological sciences. Psychology; Genetics; Hot Temperature; Microbiology; Molecular Sequence Data; N-Glycosyl Hydrolases - chemistry; N-Glycosyl Hydrolases - metabolism; Ogg1 protein; Protein Denaturation; Sequence Homology, Amino Acid; Substrate Specificity; 8-Oxoguanine DNA glycosylase; DNA repair; Archaeoglobus fulgidus; Apurinic/apyrimidinic (AP) lyase; Purification; Enzyme; Archaeobacteria; DNA-(apurinic or apyrimidinic site) lyase; Lyases; Thermal stability; Enzymatic activity; Hyperthermophily; DNA; Substrate specificity; Bacteria; Repair; Carbon-oxygen lyases; Physicochemical properties
• ISSN: 0921-8777; 0027-5107; 1386-1476
• DOI: 10.1016/S0921-8777(01)00081-7

Oxidative DNA damage is caused by reactive oxygen species formed in cells as by products of aerobic metabolism or of oxidative stress. The 8-oxoguanine (8-oxoG) DNA glycosylase from Archaeoglobus fulgidus (Afogg), which excises an oxidatively-damaged form of guanine, was overproduced in Escherichia coli, purified and characterized. A. fulgidus is a sulfate-reducing archaeon, which grows at between 60 and 95°C, with an optimum growth at 83°C. The Afogg enzyme has both DNA glycosylase and apurinic/apyrimidinic (AP) lyase activities, with the latter proceeding through a Schiff base intermediate. As expected for a protein from a hyperthermophilic organism, the enzyme activity is optimal near pH 8.5 and 60°C, denaturing at 80°C, and is thermally stable at high levels of salt (500mM). The Afogg protein efficiently cleaves oligomers containing 8-oxoG:C and 8-oxoG:G base pairs, and is less effective on oligomers containing 8-oxoG:T and 8-oxoG:A mispairs. While the catalytic action mechanism of Afogg protein is likely similar to the human Ogg1 (hOgg1), the DNA recognition mechanism and the basis for 8-oxoG substrate specificity of Afogg differ from that of hOgg.