Combinatorial targeting and discovery of ligand-receptors in organelles of mammalian cells

Nat Commun. 2012 Apr 17:3:788. doi: 10.1038/ncomms1773.

Abstract

Phage display screening allows the study of functional protein-protein interactions at the cell surface, but investigating intracellular organelles remains a challenge. Here we introduce internalizing-phage libraries to identify clones that enter mammalian cells through a receptor-independent mechanism and target-specific organelles as a tool to select ligand peptides and identify their intracellular receptors. We demonstrate that penetratin, an antennapedia-derived peptide, can be displayed on the phage envelope and mediate receptor-independent uptake of internalizing phage into cells. We also show that an internalizing-phage construct displaying an established mitochondria-specific localization signal targets mitochondria, and that an internalizing-phage random peptide library selects for peptide motifs that localize to different intracellular compartments. As a proof-of-concept, we demonstrate that one such peptide, if chemically fused to penetratin, is internalized receptor-independently, localizes to mitochondria, and promotes cell death. This combinatorial platform technology has potential applications in cell biology and drug development.

Publication types

  • Evaluation Study
  • 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
  • Bacteriophages / genetics
  • Bacteriophages / metabolism
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cell Line
  • Cell Membrane / genetics
  • Cell Membrane / metabolism
  • Cell Membrane / virology
  • Cell-Penetrating Peptides
  • Gene Expression
  • Genetic Techniques*
  • Humans
  • Ligands
  • Molecular Sequence Data
  • Organelles / genetics
  • Organelles / metabolism*
  • Protein Binding
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*

Substances

  • Carrier Proteins
  • Cell-Penetrating Peptides
  • Ligands
  • Receptors, Cell Surface
  • penetratin