TY - JOUR
T1 - Repair activities of 8-oxoguanine DNA glycosylase from Archaeoglobus fulgidus, a hyperthermophilic archaeon
AU - Chung, Ji Hyung
AU - Suh, Moo Jin
AU - Park, Young In
AU - Tainer, John A.
AU - Han, Ye Sun
N1 - Funding Information:
We are grateful to Sun-Yong Kim for assistance and valuable comments. We thank Dr. Clifford D. M. of The Scripps Research Institute for advice and a critical reading of the manuscript. We greatly thank Prof. Hiroaki Terato at Hiroshima University, Japan, for the generous gift of E. coli Fpg protein. This work was supported by International Coorperation Research Program of the Ministry of Science and Technology.
PY - 2001/7/12
Y1 - 2001/7/12
N2 - 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.
AB - 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.
KW - 8-Oxoguanine DNA glycosylase
KW - Apurinic/apyrimidinic (AP) lyase
KW - Archaeoglobus fulgidus
KW - DNA repair
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U2 - 10.1016/S0921-8777(01)00081-7
DO - 10.1016/S0921-8777(01)00081-7
M3 - Article
C2 - 11425515
AN - SCOPUS:0035850219
SN - 0921-8777
VL - 486
SP - 99
EP - 111
JO - Mutation Research - DNA Repair
JF - Mutation Research - DNA Repair
IS - 2
ER -