Native polyacrylamide gel electrophoresis (PAGE) is an invaluable technique in biochemistry to characterize native protein complexes with high molecular mass. Thus, native PAGE is suited to resolve proteasomes, giant proteases responsible for the degradation of polyubiquitylated proteins. Proteasomes contain multiple subunits and exist in different configurations. All configurations have a common 20S core particle (CP). The CP encloses the proteolytic chamber and is composed of four stacked rings with C2 symmetry. The entrance to the CP is gated by central pores within the outer rings, which also provide the binding sites for the 19S regulatory complex (RP). Adjacent regulatory proteins, such as Blm10/PA200, are bound to specific proteasome species of low abundance and contribute to the heterogeneity of proteasome complexes. To get insight into the complexity of proteasome configurations in yeast, we developed a native PAGE system by which GFP-labelled variants of proteasomal subunits are visualized by phosphofluoroimaging. Following native PAGE, proteasome species can be subjected to in-gel activity assays, subsequent SDS-PAGE, and Western blotting.
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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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Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.
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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