An Engineered Protein Antagonist of K-Ras/B-Raf Interaction

Sci Rep. 2017 Jul 19;7(1):5831. doi: 10.1038/s41598-017-05889-7.

Abstract

Ras is at the hub of signal transduction pathways controlling cell proliferation and survival. Its mutants, present in about 30% of human cancers, are major drivers of oncogenesis and render tumors unresponsive to standard therapies. Here we report the engineering of a protein scaffold for preferential binding to K-Ras G12D. This is the first reported inhibitor to achieve nanomolar affinity while exhibiting specificity for mutant over wild type (WT) K-Ras. Crystal structures of the protein R11.1.6 in complex with K-Ras WT and K-Ras G12D offer insight into the structural basis for specificity, highlighting differences in the switch I conformation as the major defining element in the higher affinity interaction. R11.1.6 directly blocks interaction with Raf and reduces signaling through the Raf/MEK/ERK pathway. Our results support greater consideration of the state of switch I and provide a novel tool to study Ras biology. Most importantly, this work makes an unprecedented contribution to Ras research in inhibitor development strategy by revealing details of a targetable binding surface. Unlike the polar interfaces found for Ras/effector interactions, the K-Ras/R11.1.6 complex reveals an extensive hydrophobic interface that can serve as a template to advance the development of high affinity, non-covalent inhibitors of K-Ras oncogenic mutants.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • HEK293 Cells
  • Humans
  • Mutant Proteins / metabolism
  • Mutation / genetics
  • Protein Binding
  • Protein Engineering*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / pharmacology*
  • ras Proteins / antagonists & inhibitors*
  • ras Proteins / metabolism

Substances

  • Mutant Proteins
  • Recombinant Proteins
  • ras Proteins