Membrane-permeable arginine-rich peptides and the translocation mechanisms

doi:10.1016/j.addr.2004.10.009

Advanced Drug Delivery Reviews

Volume 57, Issue 4, 28 February 2005, Pages 547-558

https://doi.org/10.1016/j.addr.2004.10.009 Get rights and content

Abstract

The intracellular delivery of proteins and other bioactive molecules using membrane-permeable carrier peptide vectors opens the possibility of establishing novel methods of elucidating and controlling cell functions with therapeutic potentials. One of the most typical peptide vectors is a short, arginine-rich peptide segment derived from the human immunodeficiency virus (HIV)-1 Tat protein. We have shown that not only the Tat peptide, but also various arginine-rich oligopeptides possess very similar characteristics in translocation and abilities as a delivery vector. This review summarizes the structures of these peptide vectors, especially the Tat and other arginine-rich peptides, and the current understanding of their internalization mechanisms.

Introduction

There are many bioactive molecules, the physicochemical properties of which would not allow them to cross biological membranes. Intracellular delivery of proteins involving signal transduction, for example, may regulate cellular functions. Therefore, a methodology permitting exogenous proteins into cells has become a powerful tool for cell biological studies to probe the roles of the administered proteins in the cells. When these signals are related with the occurrence or prevention of physical disorders or diseases, the introduction of such proteins can have therapeutic potential. Generally speaking, proteins have a rather hydrophilic nature and high molecular weights, which hamper their membrane permeation.

Recently, a novel strategy has been presented to use membrane-permeable peptide carrier vectors to deliver proteins into cells [1], [2], [3], [4], [5]. Short peptide segments derived from HIV-1 Tat [6] and Drosophila Antennapedia homeodomain proteins [7] are among the representatives of these peptides. By conjugation of these peptides either chemically or genetically to cargo proteins, the successful delivery of various proteins into cells with the aim of controlling cellular functions has been achieved. No significant damage to the plasma membranes and little toxicity to the cells are observed during the treatment of the cells with the peptides [8]. This strategy has been applied not only for protein delivery, but also for the delivery of various molecules with wide ranges of molecular sizes and physicochemical properties, such as small molecular weight compounds, oligonucleotides, magnetic beads, and even liposomes [1], [2], [3], [4], [5]. These characteristics may open new avenues to the establishment of novel concepts of drug delivery and therapy. Besides the high efficiency in translocation through biological membranes, the mechanisms of translocation have also become important, and considerable efforts have focused on the elucidation of the mechanisms.

In this review, we survey the structural and functional characteristics of the peptides that have been reported to have translocation activity, especially of those rich in arginines. We also shed light on the studies for the elucidation of the translocation mechanisms of these peptides, as well as their conjugates with cargoes.

Section snippets

Structural characteristics of membrane-permeable carrier peptides

Although there are various peptides that have been reported to have the similar carrier abilities, these peptides may be categorized into several classes (Table 1). Tat [6] and the oligoarginine peptides [9], [10] have a basic structure, are rich in arginine, and contain few hydrophobic amino acids. Penetratin [4], [7] was derived from the third helix of the Antennapedia homeodomain protein (positions 43–58). This peptide also contains basic amino acids for DNA binding, but is rather rich in

Tat peptide

The HIV-1 Tat is a protein composed of 86 amino acids, which binds to the trans-acting response element (TAR) of the viral RNA to transactivate the viral promoter (Tat: transactivator of HIV transcription) [24]. Since the late 1980s, it has been known that this protein is internalized into cells after addition of the protein to the culture medium [25], [26]. In 1994, Fawell et al. reported that chemical conjugation of the peptide segments derived from the Tat protein (residues 1–72 and 37–72)

Structure and internalization mechanisms of arginine-rich peptides

Besides its ability to enter cells highly efficiently even when carrying other exogenous proteins and macromolecules, HIV-1 Tat-(48–60) has been attracting our attention because of its unique characteristics in translocation. In the early days, the following properties were suggested to be cognate to the Tat peptide: (i) an ability to act as a carrier for exogenous proteins and various membrane-impermeable molecules, (ii) a rapid internalization within a few minutes, (iii) a high extent of

Conclusion

We have reviewed the structural variety of carrier peptide vectors for intracellular molecular delivery and shown that not only the Tat peptide, but also many arginine-rich peptides can be carrier vectors. A guanidino moiety in the vector seems to play a crucial role and the peptide/non-peptide carrier vectors containing guanidino functions have been introduced. These vectors promote the effective and facile delivery of proteins and other molecules that were originally unable to translocate

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Membrane-permeable arginine-rich peptides and the translocation mechanisms

Abstract

Introduction

Section snippets

Structural characteristics of membrane-permeable carrier peptides

Tat peptide

Structure and internalization mechanisms of arginine-rich peptides

Conclusion

Curr. Opin. Cell Biol.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

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Cell

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Mol. Ther.

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J. Biol. Chem.

Biochem. Biophys. Res. Comm.

Membrane permeability commonly shared among arginine-rich peptides

J. Mol. Recognit.

Modulation of cellular function by TAT mediated transduction of full length proteins

Curr. Prot. Pept. Sci.

Guanidinium rich peptide transporters and drug delivery

Curr. Prot. Pept. Sci.

Cell-Penetrating Peptides: Processes and Applications

The design, synthesis, and evaluation of molecules that enable or enhance cellular uptake: peptoid molecular transporters

Proc. Natl. Acad. Sci. U. S. A.

Evidence for an amphipathicity independent cellular uptake of amphipathic cell-penetrating peptides

Eur. J. Biochem.

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FASEB J.

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