We describe the design of a novel in vitro assay to study the interaction of soluble proteins with small hydrophobic sterol ligands. The sterol molecules are incorporated in an artificial membrane system in order to mimic their arrangement found in a biomembrane. The artificial membrane setup is monitored in real time by surface plasmon spectroscopy. Binding of fluorescently labeled soluble protein is observed by optical detection with surface plasmon enhanced fluorescence spectroscopy. By application of the novel assay, we demonstrate that four different oxidized sterol molecules are specifically recognized by the yeast protein Osh5p, a presumed oxysterol binding protein. Osh5p from yeast is the first oxysterol binding protein homologue for which oxysterol binding is shown with this new technique. With the design of our novel in vitro oxysterol binding assay, we have solved the technically challenging difficulty of presenting hydrophobic ligands to hydrophilic proteins in aqueous media.
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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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