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Article

Accelerating the Discovery of Biologically Active Small Molecules Using a High-Throughput Yeast Halo Assay

Supporting Info

Nadine C. Gassner,^† Craig M. Tamble,^† Jonathan E. Bock,^† Naomi Cotton,^† Kimberly N. White,^† Karen Tenney,^† Robert P. St. Onge,^‡ Michael J. Proctor,^§ Guri Giaever,^# Corey Nislow,^# Ronald W. Davis,^§ Phillip Crews,*^† Theodore R. Holman,*^† and R. Scott Lokey*^†

Department of Chemistry and Biochemistry, University of California, Santa Cruz, California 95064, Department of Genetics, Stanford University School of Medicine, Palo Alto, California 94305, Stanford Genome Technology Center, Palo Alto, California 94304, Faculty of Pharmacy, Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 168 College Street, Toronto, Ontario, M5S3E1, Canada, and Department of Biochemistry, Stanford University School of Medicine, Palo Alto, California

J. Nat. Prod., 2007, 70 (3), pp 383–390

DOI: 10.1021/np060555t

Publication Date (Web): February 10, 2007

Dedicated to the late Dr. Kenneth L. Rinehart of the University of Illinois at Urbana−Champaign for his pioneering work on bioactive natural products.

†

University of California, Santa Cruz.

‡

Department of Genetics, Stanford University School of Medicine.

Stanford Genome Technology Center.

University of Toronto.

Department of Biochemistry, Stanford University School of Medicine.

Corresponding author. P.C., Tel: 831-459-2603. Fax: 831-459-4197. E-mail: phil@chemistry.ucsc.edu. T.R.H., Tel: 831-459-5884. Fax: 831-459-2935. E-mail: tholman@chemistry.ucsc.edu. R.S.L., Tel: 831-459-1307. Fax: 831-459-2935. E-mail: lokey@chemistry.ucsc.edu.

Abstract

The budding yeast Saccharomyces cerevisiae, a powerful model system for the study of basic eukaryotic cell biology, has been used increasingly as a screening tool for the identification of bioactive small molecules. We have developed a novel yeast toxicity screen that is easily automated and compatible with high-throughput screening robotics. The new screen is quantitative and allows inhibitory potencies to be determined, since the diffusion of the sample provides a concentration gradient and a corresponding toxicity halo. The efficacy of this new screen was illustrated by testing materials including 3104 compounds from the NCI libraries, 167 marine sponge crude extracts, and 149 crude marine-derived fungal extracts. There were 46 active compounds among the NCI set. One very active extract was selected for bioactivity-guided fractionation, resulting in the identification of crambescidin 800 as a potent antifungal agent.

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