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Abstract
Intracellular transduction of hydrophilic macromolecules has been problematic owing to the biochemical restriction imposed by lipid bilayer of the cytoplasmic membrane. Several technologies have been developed to improve the intracellular delivery of the large molecules for therapeutic purpose, including cell penetrating peptide. Cell penetrating peptides or cell permeable peptides (CPPs) were initially discovered based on the potency of certain full-length proteins or proteins to translocate across the plasma membrane. Currently, CPPs are broadly applied for intracellular delivery of biologically functional molecules in vivo and vitro, varying from small molecules, peptides, proteins, liposomes and nucleic acids. With introducing the history and characteristics of CPPs, this review will focus on the intracellular transduction mechanism and application of CPPs.
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Figure 1.
Utility of cell permeable peptide (CPP) in transduction of various cargos into the cell. In order to utilize the CPP in intracellular transduction of various cargo molecules, (A) CPP can be covalently linked to cargos by chemical synthesis. The disulfide bonds are also used for linkage between CPP and cargos. Especially, cargos of proteins are to be expressed with CPP as recombinant proteins. (B) A wide variety of cargo has been conjugated with arginine-dependent CPPs bearing lots of positive charges. Despite the mechanism of transduction across the cellular membrane is not clearly elucidated, the initial step (1) of intracellular transduction by CPP appears to involve charge-to-charge interaction between the basic amino-acids of CPP and acidic motifs on the cell membrane. The next step (2) remains to be studied, even though there have been several hypotheses with experimental evidences for explaining how the CPPs can be internalized into cell (refer to Fig. 2). Once inside the cell (3), numerous complex events and activities can be performed on the CPP and cargo such as target binding, separation of cargo from PTD, protein refolding, post-translational modification, macromolecular assembly, nuclear import/export and so on. (B) of this figure was modified from figure 1 of the reference (2).
Figure 2.
Intracellular transduction mechanisms of cell permeable peptides. Intracellular transduction of CPPs has been explained with a variety of mechanisms. These mechanisms include (A) energy-dependent pathways, based on vesicle forming, known as endocytosis, and (B) direct translocation, which is engaged with the formation of hydrophilic pores or local destabilization of the cytoplasmic membrane. This figure was modified from figure 1 of the reference (9).
Table 1.
The representative cell-penetrating peptides (CPPs).
Table 2.
Therapeutic cases in animal models of diseases by utilizing cell-penetrating peptides (CPPs).
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