Abstract
The GDP-4-keto-6-deoxy-D-mannose epimerase/reductase (GM_ER) isolated from E. coli has been overexpressed as a GST-fusion protein and purified to homogeneity. The enzyme, an NADP+(H)-binding homodimer of 70 kDa, is responsible for the production of GDP-L-fucose. GM_ER shows significant structural homology to the human erythrocyte protein FX, which is involved in blood-group glycoconjugate biosynthesis, displaying 3,5 epimerase/reductase activity on GDP-4-keto-6-deoxy-D-mannose. GM_ER has been crystallized in a trigonal crystalline form, containing one molecule per asymmetric unit, suitable for high-resolution crystallographic investigations.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Bacterial Proteins / chemistry*
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Bacterial Proteins / isolation & purification
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Carbohydrate Epimerases / chemistry*
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Carbohydrate Epimerases / isolation & purification
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Crystallization
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Crystallography, X-Ray
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Escherichia coli / enzymology*
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Escherichia coli Proteins*
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Humans
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Ketone Oxidoreductases*
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Molecular Sequence Data
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Multienzyme Complexes*
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Protein Conformation
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / isolation & purification
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Sequence Alignment
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Sequence Homology, Amino Acid
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Sugar Alcohol Dehydrogenases / chemistry*
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Sugar Alcohol Dehydrogenases / isolation & purification
Substances
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Bacterial Proteins
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Escherichia coli Proteins
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Multienzyme Complexes
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Recombinant Fusion Proteins
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wcaG protein, E coli
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Sugar Alcohol Dehydrogenases
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Ketone Oxidoreductases
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Carbohydrate Epimerases