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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
10
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pubmed:dateCreated |
1994-7-8
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pubmed:abstractText |
A recombinant human AP endonuclease, HAP1, was constructed and characterized with respect to its ability to recognize and act upon a model double-stranded 39-mer oligodeoxyribonucleotide substrate containing a strand break site with 3'-phosphoglycolate and 5'-phosphate end-group chemistries. This oligodeoxyribonucleotide substrate exactly duplicates the chemistry and configuration of a major DNA lesion produced by ionizing radiation. HAP1 was found to recognize the strand break, and catalyze the release of the 3'-phosphoglycolate as free phosphoglycolic acid. The enzyme had a Vmax of 0.1 fmole/min/pg of HAP1 protein, and a Km of 0.05 microM for the 3'-phosphoglycolate strand break lesion. The mechanism of catalysis was hydrolysis of the phosphate ester bond between the 3'-phosphoglycolate moiety and the 3'-carbon of the adjacent dGMP moiety within the oligonucleotide. The resulting DNA contained a 3'-hydroxyl which supported nucleotide incorporation by E. coli DNA polymerase I large fragment. AP endonucleolytic activity of HAP1 was examined using an analogous double-stranded 39-mer oligodeoxyribonucleotide substrate, in which the strand break site was replaced by an apyrimidinic site. The Vmax and Km for the AP endonuclease reaction were 68 fmole/min/pg of HAP1 protein and 0.23 microM, respectively.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1371347,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1375993,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1380694,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1531616,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-15398090,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1698278,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1706477,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1719477,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1719484,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1722334,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1846559,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-1852023,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-2441359,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-2473777,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-271968,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-2847555,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-3008759,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-3047011,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-3741404,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-6162481,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-6246368,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-6822504,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7516064-942051
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/APEX1 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/DNA-(Apurinic or Apyrimidinic...,
http://linkedlifedata.com/resource/pubmed/chemical/Deoxyribonuclease IV (Phage...,
http://linkedlifedata.com/resource/pubmed/chemical/Deoxyribonucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/Endodeoxyribonucleases,
http://linkedlifedata.com/resource/pubmed/chemical/Glycolates,
http://linkedlifedata.com/resource/pubmed/chemical/RNA, Messenger,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/phosphoglycolate
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pubmed:status |
MEDLINE
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pubmed:month |
May
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pubmed:issn |
0305-1048
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
25
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pubmed:volume |
22
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1866-73
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:7516064-Amino Acid Sequence,
pubmed-meshheading:7516064-Base Sequence,
pubmed-meshheading:7516064-Cloning, Molecular,
pubmed-meshheading:7516064-DNA Damage,
pubmed-meshheading:7516064-DNA Repair,
pubmed-meshheading:7516064-DNA-(Apurinic or Apyrimidinic Site) Lyase,
pubmed-meshheading:7516064-Deoxyribonuclease IV (Phage T4-Induced),
pubmed-meshheading:7516064-Deoxyribonucleotides,
pubmed-meshheading:7516064-Endodeoxyribonucleases,
pubmed-meshheading:7516064-Glycolates,
pubmed-meshheading:7516064-HeLa Cells,
pubmed-meshheading:7516064-Humans,
pubmed-meshheading:7516064-Hydrolysis,
pubmed-meshheading:7516064-Kinetics,
pubmed-meshheading:7516064-Molecular Sequence Data,
pubmed-meshheading:7516064-RNA, Messenger,
pubmed-meshheading:7516064-Recombinant Proteins
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pubmed:year |
1994
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pubmed:articleTitle |
Removal of 3'-phosphoglycolate from DNA strand-break damage in an oligonucleotide substrate by recombinant human apurinic/apyrimidinic endonuclease 1.
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pubmed:affiliation |
Department of Radiation Medicine, Vincent T. Lombardi Cancer Research Center, Georgetown University Medical Center, Washington, DC 20007.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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