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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
41
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pubmed:dateCreated |
2004-10-6
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pubmed:abstractText |
dUTPase is essential to keep uracil out of DNA. Crystal structures of substrate (dUTP and alpha,beta-imino-dUTP) and product complexes of wild type and mutant dUTPases were determined to reveal how an enzyme responsible for DNA integrity functions. A kinetic analysis of wild type and mutant dUTPases was performed to obtain relevant mechanistic information in solution. Substrate hydrolysis is shown to be initiated via in-line nucleophile attack of a water molecule oriented by an activating conserved aspartate residue. Substrate binding in a catalytically competent conformation is achieved by (i) multiple interactions of the triphosphate moiety with catalysis-assisting Mg2+, (ii) a concerted motion of residues from three conserved enzyme motifs as compared with the apoenzyme, and (iii) an intricate hydrogen-bonding network that includes several water molecules in the active site. Results provide an understanding for the catalytic role of conserved residues in dUTPases.
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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 |
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0021-9258
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
8
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pubmed:volume |
279
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
42907-15
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pubmed:meshHeading |
pubmed-meshheading:15208312-Amino Acid Motifs,
pubmed-meshheading:15208312-Amino Acid Sequence,
pubmed-meshheading:15208312-Binding Sites,
pubmed-meshheading:15208312-Catalysis,
pubmed-meshheading:15208312-Catalytic Domain,
pubmed-meshheading:15208312-Crystallography, X-Ray,
pubmed-meshheading:15208312-DNA,
pubmed-meshheading:15208312-Electrons,
pubmed-meshheading:15208312-Escherichia coli,
pubmed-meshheading:15208312-Esters,
pubmed-meshheading:15208312-Hydrogen Bonding,
pubmed-meshheading:15208312-Hydrolysis,
pubmed-meshheading:15208312-Kinetics,
pubmed-meshheading:15208312-Ligands,
pubmed-meshheading:15208312-Magnesium,
pubmed-meshheading:15208312-Models, Chemical,
pubmed-meshheading:15208312-Models, Molecular,
pubmed-meshheading:15208312-Molecular Sequence Data,
pubmed-meshheading:15208312-Mutation,
pubmed-meshheading:15208312-Phosphates,
pubmed-meshheading:15208312-Protein Binding,
pubmed-meshheading:15208312-Pyrophosphatases,
pubmed-meshheading:15208312-Sequence Homology, Amino Acid,
pubmed-meshheading:15208312-Uracil,
pubmed-meshheading:15208312-Water
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pubmed:year |
2004
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pubmed:articleTitle |
Structural insights into the catalytic mechanism of phosphate ester hydrolysis by dUTPase.
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pubmed:affiliation |
Institute of Enzymology, Biological Research Center, Hungarian Academy of Science, Budapest, Karolina út 29-31, H-1113, Hungary.
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pubmed:publicationType |
Journal Article
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