Peptide aptamers as new tools to modulate clathrin-mediated internalisation--inhibition of MT1-MMP internalisation

BMC Cell Biol. 2010 Jul 23:11:58. doi: 10.1186/1471-2121-11-58.

Abstract

Background: Peptide aptamers are combinatorial protein reagents that bind to targets with a high specificity and a strong affinity thus providing a molecular tool kit for modulating the function of their targets in vivo.

Results: Here we report the isolation of a peptide aptamer named swiggle that interacts with the very short (21 amino acid long) intracellular domain of membrane type 1-metalloproteinase (MT1-MMP), a key cell surface protease involved in numerous and crucial physiological and pathological cellular events. Expression of swiggle in mammalian cells was found to increase the cell surface expression of MT1-MMP by impairing its internalisation. Swiggle interacts with the LLY573 internalisation motif of MT1-MMP intracellular domain, thus disrupting the interaction with the mu2 subunit of the AP-2 internalisation complex required for endocytosis of the protease. Interestingly, swiggle-mediated inhibition of MT1-MMP clathrin-mediated internalisation was also found to promote MT1-MMP-mediated cell migration.

Conclusions: Taken together, our results provide further evidence that peptide aptamers can be used to dissect molecular events mediated by individual protein domains, in contrast to the pleiotropic effects of RNA interference techniques.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Protein Complex 2 / metabolism*
  • Amino Acid Motifs
  • Aptamers, Peptide / chemistry
  • Aptamers, Peptide / isolation & purification
  • Aptamers, Peptide / pharmacology*
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cells, Cultured
  • Clathrin / metabolism
  • Endocytosis / drug effects
  • Humans
  • Matrix Metalloproteinase 14 / metabolism*
  • Protein Binding / drug effects
  • Protein Structure, Tertiary
  • SELEX Aptamer Technique

Substances

  • Adaptor Protein Complex 2
  • Aptamers, Peptide
  • Clathrin
  • Matrix Metalloproteinase 14