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. 2013 Apr;23(4):581-4.
doi: 10.1038/cr.2013.30. Epub 2013 Feb 26.

Design of stapled α-helical peptides to specifically activate Wnt/β-catenin signaling

Hong-Kui Cui, Bing Zhao, Yehua Li, Ye Guo, Hao Hu, Lei Liu, Ye-Guang Chen

Design of stapled α-helical peptides to specifically activate Wnt/β-catenin signaling

Hong-Kui Cui et al. Cell Res. 2013 Apr.
No abstract available

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Figures

Figure 1
Figure 1
Design of stapled α-helical peptides to activate Wnt/β-catenin signaling. (A) The crystal structure of the β-catenin-Axin complex with highlights of Axin (469-482) (green) and the shallow groove of β-catenin (purple and blue). (B) Schematic of peptide stapling. Two non-natural alkenyl amino acids (R8 and S5) are incorporated at two positions in the peptide chain and then cross-linked by ring-closing olefin metathesis. (C) Sequences of Axin (469-482)-derived SAHPAs. (D) Biotinylated-peptides-bound streptavidin beads were employed to pull down bacterially-expressed recombinant GST-β-catenin (133-665) fragment. The precipitants were immunoblotted with anti-GST antibody, and GST-β-catenin (133-665) inputs were shown by Coomassie staining. (E) Coimmunoprecipitation of endogenous β-catenin with Axin2 in the presence of SAHPAs (40 μM). (F) HEK293T cells were treated with Wnt3a conditioned medium (CM) and SAHPAs (40 μM) for 12 h and then harvested for immunoblotting. (G) HEK293T cells transfected with Topflash-luciferase were treated with Wnt3a CM and SAHPAs at the indicated concentrations for 12 h and then harvested for luciferase measurement. (H) Expression analyses of Wnt/β-catenin target genes by quantitative RT-PCR in HEK293T cells treated with Wnt3a CM and SAHPAs (40 μM) for 12 h. (I) R1 mESCs were cultured in N2B27 medium supplemented with LIF (10 ng/ml), BMP4 (10 ng/ml), Wnt3a (10 ng/ml), or SAHPA1 (40 μM) as indicated for 5 days and subjected to AP staining. Scale bars: 50 μm. (J) R1 cells were cultured in the conditions as described in I before being harvested for qRT-PCR. The data in G, H and J represent mean ± SD (n = 3). *** indicates P < 0.001.
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