Coupling to polymeric scaffolds stabilizes biofunctional peptides for intracellular applications

Mol Pharmacol. 2011 Apr;79(4):692-700. doi: 10.1124/mol.110.068296. Epub 2011 Jan 19.

Abstract

Here, we demonstrate that coupling to N-hydroxypropyl methacrylamide (HPMA) copolymer greatly enhances the activity of apoptosis-inducing peptides inside cells. Peptides corresponding to the BH3 domain of Bid were coupled to a thioester-activated HPMA (28.5 kDa) via native chemical ligation in a simple one-pot synthesis. Peptides and polymer conjugates were introduced into cells either by electroporation or by conjugation to the cell-penetrating peptide nona-arginine. The molecular basis of the increased activity is elucidated in detail. Loading efficiency and intracellular residence time were assessed by confocal microscopy. Fluorescence correlation spectroscopy was used as a separation-free analytical technique to determine proteolytic degradation in crude cell lysates. HPMA conjugation strongly increased the half-life of the peptides in crude cell lysates and inside cells, revealing proteolytic protection as the basis for higher activity.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / physiology
  • HeLa Cells
  • Humans
  • Intracellular Space / chemistry
  • Intracellular Space / metabolism*
  • Intracellular Space / physiology
  • Jurkat Cells
  • Methacrylates / chemistry
  • Methacrylates / metabolism*
  • Oligopeptides / chemistry
  • Oligopeptides / metabolism
  • Oligopeptides / physiology
  • Peptides / chemistry
  • Peptides / metabolism*
  • Peptides / physiology
  • Polymers / chemistry
  • Polymers / metabolism
  • Protein Binding / physiology
  • Protein Stability
  • Protein Structure, Tertiary / physiology

Substances

  • Methacrylates
  • Oligopeptides
  • Peptides
  • Polymers
  • nonaarginine
  • hydroxypropyl methacrylate