Diversity-Oriented Stapling Yields Intrinsically Cell-Penetrant Inducers of Autophagy

Leila Peraro, Zhongju Zou, Kamlesh M. Makwana, Ashleigh E. Cummings, Haydn L. Ball, Hongtao Yu, Yu Shan Lin, Beth Levine, Joshua A. Kritzer

Research output: Contribution to journalArticlepeer-review

109 Scopus citations


Autophagy is an essential pathway by which cellular and foreign material are degraded and recycled in eukaryotic cells. Induction of autophagy is a promising approach for treating diverse human diseases, including neurodegenerative disorders and infectious diseases. Here, we report the use of a diversity-oriented stapling approach to produce autophagy-inducing peptides that are intrinsically cell-penetrant. These peptides induce autophagy at micromolar concentrations in vitro, have aggregate-clearing activity in a cellular model of Huntington's disease, and induce autophagy in vivo. Unexpectedly, the solution structure of the most potent stapled peptide, DD5-o, revealed an α-helical conformation in methanol, stabilized by an unusual (i,i+3) staple which cross-links two d-amino acids. We also developed a novel assay for cell penetration that reports exclusively on cytosolic access and used it to quantitatively compare the cell penetration of DD5-o and other autophagy-inducing peptides. These new, cell-penetrant autophagy inducers and their molecular details are critical advances in the effort to understand and control autophagy. More broadly, diversity-oriented stapling may provide a promising alternative to polycationic sequences as a means for rendering peptides more cell-penetrant.

Original languageEnglish (US)
Pages (from-to)7792-7802
Number of pages11
JournalJournal of the American Chemical Society
Issue number23
StatePublished - Jun 14 2017

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry


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