Uridine diphosphoglucose 4-epimerase (EC 5.1.3.2) of Saccharomyces cerevisiae was purified to homogeneity with a yield of 30%. The purification procedure involved ammonium sulfate precipitation, streptomycin treatment, chromatography on diethylaminoethyl cellulose and hydroxylapatite, and Bio-Gel A-0.5m gel filtration. With the purified enzyme preparation, Km and Vmax values for uridine diphosphogalactose were determined and found to be 0.22 mM and 1.26 mmol/h/mg of protein, respectively. The value of Vmax corresponds to a turnover rate of 3890 molecules of uridine diphosphogalactose converted to uridine diphosphoglucose/min/enzyme molecule. The pH optimum of the enzyme was found to be between 6.8 and 8.0. Amino acid analysis was carried out on the final preparation. Based on the result, the partial specific volume was calculated to be 0.74 ml/g. The NH2-terminal residue of the enzyme was studied by two different methods and found to be threonine. The molecular weight and subunit composition were determined by the combination of the sucrose density gradient centrifugation and gel filtration under nondissociating conditions, and by polyacrylamide gel electrophoresis under dissociating conditions. The results indicated that the enzyme has a molecular weight of 183,000, consisting of two identical subunits. Each molecule of the native enzyme contained 1 molecule of NAD+.
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Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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Site | Modification | Modifier | Source | Reference |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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