Design of proteolytically stable RI-anchoring disruptor peptidomimetics for in vivo studies of anchored type I protein kinase A-mediated signalling

Biochem J. 2009 Oct 23;424(1):69-78. doi: 10.1042/BJ20090933.

Abstract

We have reported previously the design of a RIAD (RI-anchoring disruptor) peptide that specifically displaces PKA (protein kinase A) type I from the AKAP (A-kinase-anchoring protein) ezrin, which is present in the immunological synapse of T-cells. This increases immune reactivity by reducing the threshold for activation and may prove a feasible approach for improving immune function in patients with cAMP-mediated T-cell dysfunction. However, the use of RIAD in biological systems is restricted by its susceptibility to enzymatic cleavage and, consequently, its short half-life in presence of the ubiquitous serum peptidases. In the present study, carefully selected non-natural amino acids were employed in the design of RIAD analogues with improved stability. The resulting peptidomimetics demonstrated up to 50-fold increased half-lives in serum compared with RIAD, while maintaining similar or improved specificity and potency with respect to disruption of PKA type I-AKAP interactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • A Kinase Anchor Proteins / metabolism*
  • Animals
  • CSK Tyrosine-Protein Kinase
  • Cattle
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Cyclic AMP / metabolism
  • Humans
  • Interleukin-2 / metabolism
  • Mice
  • Peptides / chemical synthesis*
  • Peptides / chemistry
  • Peptides / metabolism*
  • Protein Binding
  • Protein-Tyrosine Kinases / metabolism
  • Signal Transduction / physiology*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • src-Family Kinases

Substances

  • A Kinase Anchor Proteins
  • Interleukin-2
  • Peptides
  • Cyclic AMP
  • Protein-Tyrosine Kinases
  • CSK Tyrosine-Protein Kinase
  • src-Family Kinases
  • CSK protein, human