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High-throughput screening

...And fashionable screening

Nature Reviews Drug Discovery volume 1, pages 6–7 (2002)Cite this article

Cell-based HTS systems typically express a chimeric protein consisting of the ligand-binding domain of interest coupled to a reporter. Ideally, this protein sensor should allow one ligand-binding domain to be substituted for another while always maintaining a tight association between the ligand-binding domain and the reporter, so that changes in the binding domain will be efficiently detected. Although yeast-based systems for HTS have already been described, they do not meet these requirements.

The researchers coupled a ligand-binding domain to a temperature-sensitive essential yeast metabolic enzyme, dihydrofolate reductase (DHFR), to generate a reporter in which binding to the domain resulted in yeast growth. The targets they chose to study were FKBP12 (which binds an antimicrobial agent) and the oestrogen receptor (ERα). At elevated temperatures, the transformed yeast does not grow well because the DHFR enzyme has to function with a large destabilizing insertion. However, in the presence of the specific ligands that bound to FKBP12 or ERα, DHFR had improved function and yeast growth could be detected. The yeast growth rate was also dependent on the dose of the ligand and its affinity for the receptor.

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References

ORIGINAL RESEARCH PAPER

  1. Tucker, C. L. & Fields, F. A yeast sensor of ligand binding. Nature Biotechnol. 19, 1042–1046 (2001)

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FURTHER READING

  1. Klein, C. et al. Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast. Nature Biotechnol. 16, 1334–1337 (1998)

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  2. Licitra, E. J. & Liu, J. O. A three-hybrid system for detecting small ligand–protein receptor interactions. Proc. Natl Acad. Sci. USA. 93, 12817–12821 (1996)

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Authors
  1. Melanie Brazil
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Brazil, M. ...And fashionable screening. Nat Rev Drug Discov 1, 6–7 (2002). https://doi.org/10.1038/nrd701

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