PMID- 10497264
OWN - NLM
STAT- MEDLINE
DCOM- 19991217
LR  - 20190501
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Linking)
VI  - 27
IP  - 20
DP  - 1999 Oct 15
TI  - Excision of oxidatively damaged DNA bases by the human alpha-hOgg1 protein and
      the polymorphic alpha-hOgg1(Ser326Cys) protein which is frequently found in human
      populations.
PG  - 4001-7
AB  - We have investigated the substrate specificity of the major nuclear form of the
      human Ogg1 protein, referred as alpha-hOgg1, for excision of damaged bases from
      DNA exposed to gamma-irradiation. Excision products were identified and
      quantified using gas chromatography/isotope dilution mass spectrometry (GC/IDMS).
      The GST-alpha-hOgg1 protein used in this study is a fusion of alpha-hOgg1 to the 
      C-terminus of the GST protein. The results show that GST-alpha-hOgg1 protein
      excises 8-hydroxyguanine (8-OH-Gua) and
      2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua) from DNA exposed to
      gamma-irradiation in a solution saturated with N(2)O or air. Fourteen other
      lesions, including oxidised purines and pyrimidines, were not excised from these 
      substrates. Catalytic constants were measured for the excision of 8-OH-Gua and
      FapyGua from DNA gamma-irradiated under N(2)O. The k (cat)/ K (m)values for
      excision of 8-OH-Gua and FapyGua were 4.47 x 10(-5)and 8.97 x
      10(-5)(min(-1)nM(-1)), respectively. The substrate specificity and the catalytic 
      parameters of the wild-type GST-alpha-hOgg1 protein were compared to that of a
      polymorphic form of alpha-hOgg1 harbouring a Ser-->Cys mutation at codon 326. In 
      the Japanese population, 47.6% of individuals possess both alleles coding for the
      wild-type alpha-hOgg1-Ser(326)and mutant alpha-hOgg1-Cys(326)proteins. The
      GST-alpha-hOgg1-Cys(326)protein was purified and its substrate specificity was
      determined by GC/IDMS analysis. The results show that the
      GST-alpha-hOgg1-Cys(326)protein efficiently excises 8-OH-Gua and FapyGua from
      gamma-irradiated DNA. The k (cat)/ K (m)values for excision of 8-OH-Gua and
      FapyGua were 2. 82 x 10(-5)and 4.43 x 10(-5)(min(-1)nM(-1)), respectively.
      Furthermore, we compared the capacity of these two forms of alpha-hOgg1 to act on
      substrates containing 2,6-diamino-4-hydroxy-5- N -methylformamidopyrimidine
      (Me-FapyGua). The k (cat)/ K (m)values for excision of Me-FapyGua were 278 x
      10(-5)and 319 x 10(-5)(min(-1)nM(-1)), respectively. Cleavage of 34mer
      oligodeoxyribonucleotides containing 8-OH-Gua, 8-hydroxyadenine or an
      apurinic/apyrimidinic site paired with a cytosine was also investigated. The
      results show that both GST-alpha-hOgg1-Ser(326)and
      GST-alpha-hOgg1-Cys(326)catalyse the various cleavage reactions at very similar
      rates. Furthermore, both proteins efficiently complement the mutator phenotype of
      the fpg mutY mutant of Escherichia coli.
FAU - Dherin, C
AU  - Dherin C
AD  - CEA, DSV, DRR, UMR217 CNRS-CEA, Radiobiologie Moleculaire et Cellulaire, Fontenay
      aux Roses, France.
FAU - Radicella, J P
AU  - Radicella JP
FAU - Dizdaroglu, M
AU  - Dizdaroglu M
FAU - Boiteux, S
AU  - Boiteux S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Escherichia coli Proteins)
RN  - 452VLY9402 (Serine)
RN  - EC 2.5.1.18 (Glutathione Transferase)
RN  - EC 3.2.2.- (N-Glycosyl Hydrolases)
RN  - EC 3.2.2.23 (DNA-Formamidopyrimidine Glycosylase)
RN  - EC 3.2.2.23 (DNA-formamidopyrimidine glycosylase, E coli)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Amino Acid Substitution
MH  - Cysteine/chemistry/metabolism
MH  - *DNA Damage
MH  - *DNA Repair
MH  - DNA-Formamidopyrimidine Glycosylase
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Escherichia coli
MH  - *Escherichia coli Proteins
MH  - Gas Chromatography-Mass Spectrometry
MH  - Glutathione Transferase/metabolism
MH  - Humans
MH  - N-Glycosyl Hydrolases/chemistry/*metabolism
MH  - Oxidative Stress
MH  - Phenotype
MH  - Serine/chemistry/metabolism
MH  - Substrate Specificity
PMC - PMC148667
EDAT- 1999/09/25 00:00
MHDA- 1999/09/25 00:01
CRDT- 1999/09/25 00:00
PHST- 1999/09/25 00:00 [pubmed]
PHST- 1999/09/25 00:01 [medline]
PHST- 1999/09/25 00:00 [entrez]
AID - gkc599 [pii]
AID - 10.1093/nar/27.20.4001 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 1999 Oct 15;27(20):4001-7. doi: 10.1093/nar/27.20.4001.