alpha-Isopropylmalate isomerase, the second enzyme specific for leucine biosynthesis, can be purified from extracts of yeast utilizing a chromatographic procedure that allows separation of proteins in the presence of high concentrations of (NH4)2SO4. The purification procedure utilizes the stabilizing effect of glycerol and (NH4)2SO4 on the isomerase and their opposing effects on protein retention on valine-Sepharose and leucine-Sepharose. The method effectively separates the isomerase from fumarase, a stable internal marker protein that was co-purified in early steps. High ionic strength hydrophobic chromatography, based on differential retention as a function of the length of the hydrophobic sidearm and ionic strength, yields approximately 200-fold purified alpha-isopropylmalate isomerase and may be of general utility in purifying unstable enzymes requiring high ionic strength.
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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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