The leucine zipper of the yeast transcriptional factor GCN4 was grafted to the C-terminal amino acid of the maltose binding protein (MalE) by fusing the malE gene of Escherichia coli to a synthetic gene coding for the leucine zipper. The hybrid protein, MalE-Lzp, was synthesized in large amounts from multicopy plasmids and efficiently exported into the periplasmic space of E. coli, up to 200,000 molecules per cell. Unlike hybrids between MalE and other proteins, MalE-Lzp was quite stable exhibiting only minimal degradation. The hybrid was purified from a periplasmic extract in one step by affinity chromatography on cross-linked amylose. Sedimentation velocity and gel filtration experiments showed that MalE-Lzp existed as a dimer in conditions where MalE was a monomer, at all concentrations tested down to 0.1 microM. Thus, it was possible to engineer the quaternary structure of an exported, monomeric protein by using a structural motif taken from a natural protein.
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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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