Protease-Resistant and Cell-Permeable Double-Stapled Peptides Targeting the Rab8a GTPase

ACS Chem Biol. 2016 Aug 19;11(8):2375-82. doi: 10.1021/acschembio.6b00386. Epub 2016 Jul 11.

Abstract

Small GTPases comprise a family of highly relevant targets in chemical biology and medicinal chemistry research and have been considered "undruggable" due to the persisting lack of effective synthetic modulators and suitable binding pockets. As molecular switches, small GTPases control a multitude of pivotal cellular functions, and their dysregulation is associated with many human diseases such as various forms of cancer. Rab-GTPases represent the largest subfamily of small GTPases and are master regulators of vesicular transport interacting with various proteins via flat and extensive protein-protein interactions (PPIs). The only reported synthetic inhibitor of a PPI involving an activated Rab GTPase is the hydrocarbon stapled peptide StRIP3. However, this macrocyclic peptide shows low proteolytic stability and cell permeability. Here, we report the design of a bioavailable StRIP3 analogue that harbors two hydrophobic cross-links and exhibits increased binding affinity, combined with robust cellular uptake and extremely high proteolytic stability. Localization experiments reveal that this double-stapled peptide and its target protein Rab8a accumulate in the same cellular compartments. The reported approach offers a strategy for the implementation of biostability into conformationally constrained peptides while supporting cellular uptake and target affinity, thereby conveying drug-like properties.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Biological Availability
  • HeLa Cells
  • Humans
  • Peptide Hydrolases / metabolism*
  • Peptides / chemistry
  • Peptides / metabolism*
  • Permeability
  • rab GTP-Binding Proteins / metabolism*

Substances

  • Peptides
  • Peptide Hydrolases
  • RAB8A protein, human
  • rab GTP-Binding Proteins