Phage display has demonstrated the utility of cyclic peptides as general protein ligands but cannot access proteins inside eukaryotic cells. Expanding a new chemical genetics tool, we describe the first expressed library of head-to-tail cyclic peptides in yeast (Saccharomyces cerevisiae). We applied the library to selections in a yeast model of α-synuclein toxicity that recapitulates much of the cellular pathology of Parkinson's disease. From a pool of 5 million transformants, we isolated two related cyclic peptide constructs that specifically reduced the toxicity of human α-synuclein. These expressed cyclic peptide constructs also prevented dopaminergic neuron loss in an established Caenorhabditis elegans Parkinson's model. This work highlights the speed and efficiency of using libraries of expressed cyclic peptides for forward chemical genetics in cellular models of human disease.
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Filamentous Virus-based Assembly: Their Oriented Structures and Thermal Diffusivity
Scientific Reports Open Access 03 April 2018
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We thank S. Benkovic (Pennsylvania State University) for plasmids, E. Spooner for mass spectrometry assistance and N. Azubuine for preparation of media. This work was supported by a National Research Service Award fellowship from the US National Institute of Neurological Disorders and Stroke (NINDS) and National Institute on Aging (J.A.K.), an R21 grant from NINDS (S.L. and J.A.K.) and the Morris K. Udall Centers of Excellence for Parkinson's Disease Research (S.L.) Parkinson's disease research in the Caldwell Lab (G.A.C, K.A.C., S.H.) was supported by the American Parkinson Disease Association, Michael J. Fox Foundation and US National Institute of Environmental Health Sciences.
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Kritzer, J., Hamamichi, S., McCaffery, J. et al. Rapid selection of cyclic peptides that reduce α-synuclein toxicity in yeast and animal models. Nat Chem Biol 5, 655–663 (2009). https://doi.org/10.1038/nchembio.193
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