Mammalian display screening of diverse cystine-dense peptides for difficult to drug targets

• Published in: Nature communications Vol. 8; no. 1; p. 2244
• Main Authors: Crook, Zachary R., Sevilla, Gregory P., Friend, Della, Brusniak, Mi-Youn, Bandaranayake, Ashok D., Clarke, Midori, Gewe, Mesfin, Mhyre, Andrew J., Baker, David, Strong, Roland K., Bradley, Philip, Olson, James M.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 21-12-2017; Nature Publishing Group; Nature Portfolio
• Subjects: 631/154/51/2313; 631/1647/2163; 631/61/338/469; 631/67/1059/153; Amino Acid Sequence; Animals; Cystine; Cystine - analysis; Drug development; Drug Discovery; Escherichia coli Proteins - chemistry; Glycosylation; Humanities and Social Sciences; Humans; Mammals; Medical treatment; Molecular modelling; multidisciplinary; Peptide Library; Peptides; Peptides - chemistry; Peptides - metabolism; Peptides - pharmacology; Protein Binding; Protein Folding; Protein interaction; Protein Interaction Maps - drug effects; Reproducibility of Results; Saccharomyces cerevisiae Proteins - chemistry; Science; Science (multidisciplinary); Stability; Yes-associated protein
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-017-02098-8

Protein:protein interactions are among the most difficult to treat molecular mechanisms of disease pathology. Cystine-dense peptides have the potential to disrupt such interactions, and are used in drug-like roles by every clade of life, but their study has been hampered by a reputation for being difficult to produce, owing to their complex disulfide connectivity. Here we describe a platform for identifying target-binding cystine-dense peptides using mammalian surface display, capable of interrogating high quality and diverse scaffold libraries with verifiable folding and stability. We demonstrate the platform’s capabilities by identifying a cystine-dense peptide capable of inhibiting the YAP:TEAD interaction at the heart of the oncogenic Hippo pathway, and possessing the potency and stability necessary for consideration as a drug development candidate. This platform provides the opportunity to screen cystine-dense peptides with drug-like qualities against targets that are implicated for the treatment of diseases, but are poorly suited for conventional approaches.
Pathologies related to protein:protein interaction are hard to treat but cystine-dense peptides have the potential to disrupt such interactions. Here the authors develop a high-diversity mammalian cell screen for cystine-dense peptides with drug potential and use it to identify a YAP:TEAD inhibitor.