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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
32
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pubmed:dateCreated |
1993-9-16
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pubmed:abstractText |
Reductive methylation and site-directed mutagenesis experiments have implicated the N-terminal alpha-amino group of T4 endonuclease V in the glycosylase and abasic lyase activities of the enzyme. NMR studies have confirmed the involvement of the N-terminal alpha-amino group in the inhibition of enzyme activity by reductive methylation. A mechanism accounting for these results predicts that a (imino) covalent enzyme-substrate intermediate is formed between the protein N-terminal alpha-amino group and C1' of the 5'-deoxyribose of the pyrimidine dimer substrate subsequent to (or concomitantly with) the glycosylase step. Experiments to verify the existence of this intermediate indicated that enzyme inhibition by cyanide was substrate-dependent, a result classically interpreted to imply an imino reaction intermediate. In addition, sodium borohydride reduction of the intermediate formed a stable dead-end enzyme-substrate product. This product was formed whether ultraviolet light-irradiated high molecular weight DNA or duplex oligonucleotides containing a defined thymine-thymine cyclobutane dimer were used as substrate. The duplex oligonucleotide substrates demonstrated a well-defined gel shift. This will facilitate high-resolution footprinting of the enzyme on the DNA substrate.
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pubmed:grant |
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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/Borohydrides,
http://linkedlifedata.com/resource/pubmed/chemical/Cyanogen Bromide,
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/Deoxyribonuclease (Pyrimidine Dimer),
http://linkedlifedata.com/resource/pubmed/chemical/Endodeoxyribonucleases,
http://linkedlifedata.com/resource/pubmed/chemical/Imines,
http://linkedlifedata.com/resource/pubmed/chemical/Pyrimidine Dimers,
http://linkedlifedata.com/resource/pubmed/chemical/Viral Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/endonuclease V, phage T4,
http://linkedlifedata.com/resource/pubmed/chemical/sodium borohydride
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pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
0006-2960
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
17
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pubmed:volume |
32
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
8284-90
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:8347626-Animals,
pubmed-meshheading:8347626-Base Sequence,
pubmed-meshheading:8347626-Binding Sites,
pubmed-meshheading:8347626-Borohydrides,
pubmed-meshheading:8347626-Cattle,
pubmed-meshheading:8347626-Cyanogen Bromide,
pubmed-meshheading:8347626-DNA,
pubmed-meshheading:8347626-DNA Damage,
pubmed-meshheading:8347626-DNA Repair,
pubmed-meshheading:8347626-Deoxyribonuclease (Pyrimidine Dimer),
pubmed-meshheading:8347626-Endodeoxyribonucleases,
pubmed-meshheading:8347626-Imines,
pubmed-meshheading:8347626-Magnetic Resonance Spectroscopy,
pubmed-meshheading:8347626-Methylation,
pubmed-meshheading:8347626-Molecular Sequence Data,
pubmed-meshheading:8347626-Mutagenesis, Site-Directed,
pubmed-meshheading:8347626-Pyrimidine Dimers,
pubmed-meshheading:8347626-Structure-Activity Relationship,
pubmed-meshheading:8347626-Ultraviolet Rays,
pubmed-meshheading:8347626-Viral Proteins
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pubmed:year |
1993
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pubmed:articleTitle |
Evidence for an imino intermediate in the T4 endonuclease V reaction.
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pubmed:affiliation |
Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston 77555.
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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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