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Total Chemical Synthesis and Folding of All-l and All-d Variants of Oncogenic KRas(G12V)

Adam M Levinson1, John H McGee2, Andrew G Roberts

  • 1Tri-Institutional PhD Program in Chemical Biology, Weill Cornell Medical College , New York, New York 10065, United States.

Journal of the American Chemical Society
|April 28, 2017
PubMed

Insights

Researchers synthesized mirror-image forms of cancer-driving Ras proteins (KRas). These enantiomeric proteins can fold and bind molecules, enabling new therapeutic strategies against difficult cancer targets.

Area of Science:

  • Biochemistry
  • Chemical Biology
  • Oncology

Background:

  • Ras proteins are crucial GTPases regulating cell growth and survival.
  • Mutated Ras proteins are implicated in ~30% of human cancers, driving uncontrolled proliferation.
  • Ras has been historically challenging to target therapeutically due to its biochemical properties.

Purpose of the Study:

  • To achieve total chemical synthesis of all-l- and all-d-amino acid variants of oncogenic KRas(G12V).
  • To investigate the biochemical properties and functional competency of these enantiomeric Ras proteins.
  • To establish a foundation for developing novel therapeutic strategies against oncogenic Ras.

Main Methods:

  • Fmoc-based solid-phase peptide synthesis was employed for protein construction.
  • Native chemical ligation and isonitrile-mediated activation were used for assembly.
  • Enantiomeric KRas(G12V) variants were synthesized and biochemically characterized.

Main Results:

  • Successfully synthesized all-l- and all-d-amino acid biotinylated KRas(G12V) variants.
  • Demonstrated that both enantiomers of KRas(G12V) can fold and bind nucleotide substrates.
  • Observed enantiodiscrimination in nucleotide substrate and binding partner interactions.

Conclusions:

  • The synthesis and functional validation of enantiomeric KRas provide a novel biochemical tool.
  • This work enables mirror-image yeast surface display for identifying all-d peptide ligands.
  • These findings offer a promising new avenue for developing therapeutics against oncogenic KRas.

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