Use of a penetratin-linked peptide in Dictyostelium

Author: Ryves W.   Harwood Adrian  

Publisher: Humana Press, Inc

ISSN: 1073-6085

Source: Molecular Biotechnology, Vol.33, Iss.2, 2006-06, pp. : 123-131

Disclaimer: Any content in publications that violate the sovereignty, the constitution or regulations of the PRC is not accepted or approved by CNPIEC.

Previous Menu Next

Abstract

The plasma membrane is an effective barrier to most macromolecules and hydrophilic molecules. Remarkably, a class of positively charged cell-penetrating peptides (CPPs) has been discovered that can translocate themselves and associated cargoes into the cytoplasm. These have been used to carry oligopeptide- and oligonucleotide-based inhibitors into mammalian cells. A recent report indicates that the same CPPs are internalized by plant protoplasts, suggesting that this may be a universal phenomenon. We report here that the prototypical CPP, penetratin, enters cells of the free-living amoebae Dictyostelium discoideum. To investigate the functionality of this technology, we fused the penetratin sequence to PKI, a peptide inhibitor of the cAMP-dependent protein kinase (PKA). Consistent with its PKA inhibitory action, Penetratin-PKI blocked aggregation in wild-type cells and, at appropriate concentrations, rescued the phenotype of a Dictyostelium mutant that has constitutively high PKA activity. This technology offers an effective method for delivery of oligopeptides and oligonucleotides into Dictyostelium.

Related content