Poster Presentation 12th Australian Peptide Conference 2017

Downsizing proto oncogene cfos to short cell permeable helix constrained peptides (#118)

Samuel R Perry 1 , Daniel Baxter 2 , Tim A Hill 1 , Woan M Kok 1 , Nathan R Zaccai 3 , Leo Brady 3 , Jody M Mason 2 , David P Fairlie 1
  1. Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia
  2. University of Bath, Bath, United Kingdom
  3. University of Bristol, Bristol, United Kingdom

The oncogenic transcription factor activator protein-1 (AP-1) is a DNA-binding protein that assembles through dimerization of leucine-rich helical sequences of Fos and Jun protein subunits into a coiled-coil. The  proto-oncogene cFos (380 residues) was downsized to shorter peptides (37-25 residues) modified with helix-inducing i, i+4 lactam (Lys1-Asp5) constraints to enhance binding to Jun. Structural integrity in the presence and absence of Jun was assessed by circular dichroism spectroscopy, while the thermodynamics of binding was measured by isothermal titration calorimetry. Substitution of the i, i+4 lactams to i, i+4 hydrocarbon staples improved cell permeability, but not to the same magnitude as addition of cationic cell penetrating peptides. Maintaining the helix inducing i, i+4 lactam constraints with the appended cell penetrating peptide, including a nuclear localisation sequence, was the most effective at promoting cell uptake and nuclear localisation.