Abstract
Cell permeating peptides (CPPs) are attracting great interest for use as molecular delivery vehicles for the transport of biologically active cargo across the cell membrane. The sequence of a novel CPP sequence, termed ‘Cupid’, was identified from the genome of Dictyostelium discoideum. A Cupid-Green Fluorescent Protein (Cupid-GFP) fusion protein was tested on mammalian, whole plant cells, plant leaf protoplast and fungal cell cultures and observed using confocal microscopy. GFP fluorescence builds up within the cell cytosol in 60 min, demonstrating Cupid-GFP has permeated them and folded correctly into its fluorescent form. Our combined data suggest Cupid can act as a molecular vehicle capable of delivering proteins, such as GFP, into the cytosol of a variety of cells.
Highlights
Cell permeating peptides (CPPs) are attracting great interest for use as molecular delivery vehicles for the transport of biologically active cargo across the cell membrane
This paper introduces a novel cell permeating peptide sequence identified in the genome of the free-living amoeba, Dictyostelium discoideum, termed ‘Cupid’, (Cellular Permeability factor in Dictyostelium)
To investigate the nature of this capability, an expression plasmid was constructed to link the carboxy-terminus of the Cupid peptide to the Aequorea victorea Green Fluorescent Protein (GFP) sequence, and the fusion protein was purified using an N terminal polyhistidine motif
Summary
Cell permeating peptides (CPPs) are attracting great interest for use as molecular delivery vehicles for the transport of biologically active cargo across the cell membrane. CPPs may be classified based upon their binding properties to lipids linked to their peptide sequences: primary amphipathic; secondary amphipathic; and non-amphipathic[11,12,13] They can be classified according to the physical mechanism of cellular uptake, for example whether it is endocytosis-mediated or direct permeation of the membrane. Macropinocytosis, clathrin-coated pits and caveolae/ lipid-raft-mediated endocytosis mechanisms have been implicated through their differing sensitivity to drug applications[14,15,16,17,18,19] These processes involve energetic membrane folding and employ internalization proteins that require cellular ATP to function.
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