Abstract
A human dihydropteridine reductase (EC 1.6.99.10) has been created from a rat cDNA clone by a single five-oligonucleotide mutagenesis reaction and expressed in good yield in Escherichia coli. The enzyme has been purified to homogeneity, and kinetic identity to the naturally occurring enzyme has been proven. Crystallization has also been achieved, and the crystal structure was solved using 2.5 A data that was refined to an R value of 16.9%. The structure described in this report represents the first complete structural characterization of this important human enzyme.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Base Sequence
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Crystallography, X-Ray
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DNA, Complementary
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Dihydropteridine Reductase / biosynthesis
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Dihydropteridine Reductase / chemistry*
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Dihydropteridine Reductase / genetics
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Escherichia coli / genetics
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Humans
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Kinetics
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Molecular Sequence Data
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Mutagenesis
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NAD / chemistry*
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Protein Conformation
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Rats
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Sequence Homology, Amino Acid
Substances
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DNA, Complementary
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Recombinant Proteins
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NAD
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Dihydropteridine Reductase