The NH2 terminus of influenza virus hemagglutinin-2 subunit peptides enhances the antitumor potency of polyarginine-mediated p53 protein transduction

J Biol Chem. 2005 Mar 4;280(9):8285-9. doi: 10.1074/jbc.M412430200. Epub 2004 Dec 16.

Abstract

Protein transduction therapy is a newly developing method that allows proteins, peptides, and biologically active compounds to penetrate across the plasma membrane by being fused with cell-penetrating peptides such as polyarginine. Polyarginine-fused p53 protein penetrates across the plasma membrane of cancer cells and inhibits the growth of the cells. However, the protein is often entrapped inside macropinosomes in the cytoplasm. Therefore, high dose concentrations of the protein are needed for it to function effectively. To overcome this problem, in the present study, polyarginine-fused p53 was linked with the NH(2)-terminal domain of influenza virus hemagglutinin-2 subunit (HA2), which is a pH-dependent fusogenic peptide that induces the lysis of membranes at low pH levels. The protein was capable of efficiently translocating into the nucleus of glioma cells and induced p21(WAF1) transcriptional activity more effectively than did polyarginine-fused p53 protein. Moreover, low concentrations of the protein significantly inhibited the growth of cancer cells. These results suggest that protein transduction therapy using polyarginine and HA2 may be useful as a method for cancer therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology*
  • Apoptosis
  • Astrocytes / drug effects
  • Astrocytes / metabolism*
  • Blotting, Western
  • Cell Cycle Proteins / metabolism
  • Cell Line, Tumor
  • Cell Membrane / virology
  • Cell Proliferation
  • Cell Survival
  • Cells, Cultured
  • Cyclin-Dependent Kinase Inhibitor p21
  • Drug Synergism*
  • Fluorescein-5-isothiocyanate / pharmacology
  • Genes, Reporter
  • Hemagglutinins, Viral / chemistry*
  • Humans
  • Hydrogen-Ion Concentration
  • In Situ Nick-End Labeling
  • Mutagenesis, Site-Directed
  • Neoplasms / drug therapy*
  • Neoplasms / virology
  • Orthomyxoviridae / metabolism*
  • Peptides / chemistry*
  • Protein Binding
  • Protein Structure, Tertiary
  • Rats
  • Rats, Wistar
  • Recombinant Fusion Proteins / chemistry
  • Time Factors
  • Transcriptional Activation
  • Tumor Suppressor Protein p53 / chemistry
  • Tumor Suppressor Protein p53 / metabolism*

Substances

  • Antineoplastic Agents
  • CDKN1A protein, human
  • Cdkn1a protein, rat
  • Cell Cycle Proteins
  • Cyclin-Dependent Kinase Inhibitor p21
  • Hemagglutinins, Viral
  • Peptides
  • Recombinant Fusion Proteins
  • Tumor Suppressor Protein p53
  • hemagglutinin HA-2 fusogenic peptide, Influenza virus
  • polyarginine
  • Fluorescein-5-isothiocyanate