TY - JOUR
T1 - Diversity-Oriented Stapling Yields Intrinsically Cell-Penetrant Inducers of Autophagy
AU - Peraro, Leila
AU - Zou, Zhongju
AU - Makwana, Kamlesh M.
AU - Cummings, Ashleigh E.
AU - Ball, Haydn L.
AU - Yu, Hongtao
AU - Lin, Yu Shan
AU - Levine, Beth
AU - Kritzer, Joshua A.
N1 - Publisher Copyright:
© 2017 American Chemical Society.
PY - 2017/6/14
Y1 - 2017/6/14
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85020813865&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85020813865&partnerID=8YFLogxK
U2 - 10.1021/jacs.7b01698
DO - 10.1021/jacs.7b01698
M3 - Article
C2 - 28414223
AN - SCOPUS:85020813865
SN - 0002-7863
VL - 139
SP - 7792
EP - 7802
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 23
ER -