GPI-APs are a family of proteins attached to the plasma membrane by a glycoplipid that undergoes remodeling of the glycan and lipid structure during transport to the cell surface. We describe a protocol to induce the synthesis of a GPI-anchored protein whereby EtNP is added to Man2 but not removed. By temporally manipulating the expression of Gpi7p, the enzyme that adds EtNP to Man2, in ted1Δ dcr2Δ cells prior to the expression of a canonical GPI-AP (mNeon-Gas1p), EtNP is attached to Man2 of de novo synthesized mNeon-Gas1p and cannot be removed. This strategy provides a means to temporally and spatially track the transport of remodeling-defective GPI-APs in yeast cells. For complete details on the use and execution of this protocol, please refer to Chen et al. (2021).
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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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Evidence ID | Analyze ID | File | Description |
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