A protein-targeting strategy used to develop a selective inhibitor of the E17K point mutation in the PH domain of Akt1

Nat Chem. 2015 May;7(5):455-62. doi: 10.1038/nchem.2223. Epub 2015 Apr 13.

Abstract

Ligands that can bind selectively to proteins with single amino-acid point mutations offer the potential to detect or treat an abnormal protein in the presence of the wild type (WT). However, it is difficult to develop a selective ligand if the point mutation is not associated with an addressable location, such as a binding pocket. Here we report an all-chemical synthetic epitope-targeting strategy that we used to discover a 5-mer peptide with selectivity for the E17K-transforming point mutation in the pleckstrin homology domain of the Akt1 oncoprotein. A fragment of Akt1 that contained the E17K mutation and an I19[propargylglycine] substitution was synthesized to form an addressable synthetic epitope. Azide-presenting peptides that clicked covalently onto this alkyne-presenting epitope were selected from a library using in situ screening. One peptide exhibits a 10:1 in vitro selectivity for the oncoprotein relative to the WT, with a similar selectivity in cells. This 5-mer peptide was expanded into a larger ligand that selectively blocks the E17K Akt1 interaction with its PIP3 (phosphatidylinositol (3,4,5)-trisphosphate) substrate.

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
  • Epitopes / chemistry
  • Ligands
  • Molecular Sequence Data
  • Point Mutation*
  • Protein Binding
  • Proto-Oncogene Proteins c-akt / chemistry
  • Proto-Oncogene Proteins c-akt / genetics
  • Proto-Oncogene Proteins c-akt / metabolism*

Substances

  • Epitopes
  • Ligands
  • AKT1 protein, human
  • Proto-Oncogene Proteins c-akt

Associated data

  • PubChem-Substance/249732336
  • PubChem-Substance/249732337
  • PubChem-Substance/249732338
  • PubChem-Substance/249732339
  • PubChem-Substance/249732340
  • PubChem-Substance/249732341
  • PubChem-Substance/249732342
  • PubChem-Substance/249732343
  • PubChem-Substance/249732344
  • PubChem-Substance/249732345
  • PubChem-Substance/249737687