A Yeast Ligand Sensor

Introduction The yeast ligand sensor is a novel approach for the detection and quantification of ligandbinding, an issue of great interest in drug discovery and other areas. The approach is a protein-derived biosensor in which the readout is a change in growth of temperaturesensitive yeast. The yeast lack dihydrofolate reductase (DHFR) and are complemented by mouse DHFR containing a ligand-binding domain inserted in a flexible loop. The yeast grow poorly and are temperature-sensitive, due in large part to the inserted ligand-binding domain. In the presence of a ligand specific to the binding domain, yeast are less temperature-sensitive and grow better, thus allowing easy detection of ligand binding. Technology description The researchers have generated 2 examples of yeast expressing DHFR ligand-binding domain fusions, FKBP-DHFR and estrogen receptor (ERα)-DHFR, and provide here the yeast strains (TH5) expressing these fusion proteins or the DNA plasmids. Business Opportunity The potential uses of this sensor include screening chemical libraries for novel compounds that bind a protein of interest, screening variants of a binding domain for mutations that affect ligand association, and selecting and evolving ligand-binding variants from pools of mutagenized domains. As with other yeast-based approaches, it has the advantages of being simple, inexpensive, and adaptable. Related Publication(s)
Nat Biotechnol. 2001 Nov;19(11):1042-6

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