Arginine-rich Peptides

Futaki, Shiroh; Suzuki, Tadanao; Ohashi, Wakana; Yagami, Takeshi; Tanaka, Seigo; Ueda, Kunihiro; Sugiura, Yukio

doi:10.1074/jbc.m007540200

Cited by 1,498 publications

(625 citation statements)

References 19 publications

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“…[41][42][43][44][45][46][47][48][49][50] On the basis of these past reports and the results of our current study, we sought to determine whether ultrathin multilayered RNase A-R 9 /SPS films could be used to promote the surface-mediated delivery of RNase A to cells.…”

Section: Surface-mediated Delivery Of Rnase A-r 9 To Cellsmentioning

confidence: 99%

Multilayered Films Fabricated from an Oligoarginine-Conjugated Protein Promote Efficient Surface-Mediated Protein Transduction

et al. 2007

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The conjugation of cationic protein transduction domains to proteins results in an increase in the extent to which proteins are internalized by cells. This investigation sought to determine whether the conjugation of a protein transduction domain to a functional protein could be used to facilitate the incorporation of the protein into multilayered polyelectrolyte films and, subsequently, whether these films could be used to promote surface-mediated protein transduction. We demonstrate that it is possible to fabricate multilayered assemblies 80 nm thick using sodium polystyrene sulfonate (SPS) and bovine pancreatic ribonuclease (RNase A) conjugated to the cationic protein transduction domain nonaarginine (R 9 ) using an entirely aqueous layer-by-layer process. We demonstrate further that the conjugation of R 9 to RNase A permits the assembly of multilayered films under conditions that do not allow for the incorporation of the unmodified protein. This result suggests that R 9 functions as a cationic anchor and serves to increase the strength of electrostatic interactions with SPS and facilitate layer-by-layer assembly. We also demonstrate that RNase A-R 9 /SPS films dissolve rapidly in physiologically relevant media and that macroscopic objects coated with these materials can be used to mediate high levels of protein transduction in mammalian cells. These results suggest the basis of general methods that could contribute to the design of materials that permit spatial and temporal control over the delivery of therapeutic proteins to cells and tissues.

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Section: Surface-mediated Delivery Of Rnase A-r 9 To Cellsmentioning

confidence: 99%

Multilayered Films Fabricated from an Oligoarginine-Conjugated Protein Promote Efficient Surface-Mediated Protein Transduction

et al. 2007

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“…Because these peptides are highly charged and therefore highly hydrophilic, their spontaneous translocation across the hydrophobic core of the membrane is difficult to understand. The most common mechanisms suggested for spontaneous translocation across cell membranes, such as the inverse micelle model (20), the carpet model (21), or the pore-formation model (22) formed by amphipathic ␣-helical peptides, do not provide a satisfactory explanation of how these peptides are able to translocate.Experimental studies show that the L and D amino acid sequences, as well as the reverse Tat sequence, can translocate across membranes, suggesting that the secondary structure of the peptide does not affect its ability to translocate (18,23). Another important finding has been that translocation requires a relatively large concentration of peptides.…”

mentioning

confidence: 97%

Molecular dynamics simulations suggest a mechanism for translocation of the HIV-1 TAT peptide across lipid membranes

Herce

Garcı́a

2007

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

399

454

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The recombinant HIV-1 Tat protein contains a small region corresponding to residues 47 YGRKKRRQRR 57 R, which is capable of translocating cargoes of different molecular sizes, such as proteins, DNA, RNA, or drugs, across the cell membrane in an apparently energy-independent manner. The pathway that these peptides follow for entry into the cell has been the subject of strong controversy for the last decade. This peptide is highly basic and hydrophilic. Therefore, a central question that any candidate mechanism has to answer is how this highly hydrophilic peptide is able to cross the hydrophobic barrier imposed by the cell membrane. We propose a mechanism for the spontaneous translocation of the Tat peptides across a lipid membrane. This mechanism involves strong interactions between the Tat peptides and the phosphate groups on both sides of the lipid bilayer, the insertion of charged side chains that nucleate the formation of a transient pore, followed by the translocation of the Tat peptides by diffusing on the pore surface. This mechanism explains how key ingredients, such as the cooperativity among the peptides, the large positive charge, and specifically the arginine amino acids, contribute to the uptake. The proposed mechanism also illustrates the importance of membrane fluctuations. Indeed, mechanisms that involve large fluctuations of the membrane structure, such as transient pores and the insertion of charged amino acid side chains, may be common and perhaps central to the functions of many membrane protein functions.cell-penetrating peptide ͉ antimicrobial peptide ͉ drug delivery ͉ membrane proteins ͉ pore formation T he HIV-1 Tat protein indicated that some proteins might have short sequence segments responsible for their translocation across cell membranes in a seemingly energy-independent manner (1-8). The sequence responsible for the cellular uptake of the HIV-1 Tat corresponds to residues 47 YGRKKRRQRR 57 R. This peptide is called the Tat peptide, and it has been categorized as a member of a family of peptides called cell-penetrating peptides. These peptides consist of short sequences (10-30 aa) that are highly basic and enter the cells in a seemingly energy-independent manner (1). This property makes them extraordinarily good candidates as transporters for drug delivery (7, 9-11). Therefore, significant effort is currently being invested to understand this phenomenon at a fundamental molecular level, as well as to employ these peptides as drug carriers (2,(12)(13)(14)(15)(16).Much debate exists around what makes it possible for these peptides to translocate across biological membranes. A diverse set of experiments indicates that there may be more than one mechanism contributing to the uptake of these peptides (1-7, 11, 17-20). Among those mechanisms is the possibility that these peptides may be able to spontaneously translocate across the cell membrane, as suggested by several experiments that indicate a nonendocytotic or energy-independent pathway for the uptake. Because these peptides are highly ...

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“…The uptake of the penetratin and the HIV-1 Tat peptide had originally been described to be insensitive to low temperature (21,22,24) and to inhibitors of endocytosis (22,25). Penetratin was also demonstrated to traverse a pure lipid bilayer (26) without forming pores (26,27).…”

mentioning

confidence: 99%

A Stepwise Dissection of the Intracellular Fate of Cationic Cell-penetrating Peptides

Fischer

Köhler

Fotin‐Mleczek

et al. 2004

Journal of Biological Chemistry

270

289

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The role of endosomal acidification and retrograde transport for the uptake of the highly basic cell-penetrating peptides penetratin, Tat, and oligoarginine was investigated. The effect of a panel of drugs that interfere with discrete steps of endocytosis or Golgi-mediated transport on uptake and cellular distribution of fluorescein-labeled peptide analogues was probed by confocal microscopy, flow cytometry, and fluorescence spectroscopy of whole cell lysates. The analyses were carried out in MC57 fibrosarcoma cells and in HeLa cells. While MC57 fibrosarcoma cells showed some vesicular fluorescence and a pronounced cytoplasmic fluorescence, in HeLa cells little cytoplasmic fluorescence was observed. In MC57 cells the inhibitors of endosomal acidification chloroquine and bafilomycin A1 abolished the release of the peptides into the cytoplasm. Release into the cytosol preserved endosomal integrity. In addition, cellular uptake of the peptides was inhibited by brefeldin A, a compound interfering with trafficking in the transGolgi network. In contrast, nordihydroguaiaretic acid, a drug that stimulates the rapid retrograde movement of both Golgi stacks and trans-Golgi network to the endoplasmic reticulum, promoted a cytoplasmic localization of Tat peptides in peptide-pulsed HeLa cells. The effects of these drugs on trafficking shared characteristics with those reported for the trafficking of plant and bacterial toxins, such as cholera toxin, which reach the cytoplasm by means of retrograde transport. A sequence comparison revealed a common stretch of 8 -10 amino acids with high sequence homology to the Tat peptide. The structural and functional data therefore strongly suggest a common mechanism of import for cationic cell-penetrating peptides and the toxins.

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Arginine-rich Peptides

Cited by 1,498 publications

References 19 publications

Multilayered Films Fabricated from an Oligoarginine-Conjugated Protein Promote Efficient Surface-Mediated Protein Transduction

Multilayered Films Fabricated from an Oligoarginine-Conjugated Protein Promote Efficient Surface-Mediated Protein Transduction

Molecular dynamics simulations suggest a mechanism for translocation of the HIV-1 TAT peptide across lipid membranes

A Stepwise Dissection of the Intracellular Fate of Cationic Cell-penetrating Peptides

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