Methodological and cellular aspects that govern the internalization mechanisms of arginine-rich cell-penetrating peptides

Nakase, Ikuhiko; Takeuchi, Toshihide; Tanaka, Gen; Futaki, Shiroh

doi:10.1016/j.addr.2007.10.006

Cited by 338 publications

(265 citation statements)

References 86 publications

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“…Although the mechanism of PTD and sc-protein uptake is still poorly defined (1,(6)(7)(8), it is clear that their extracellular concentration dictates the efficiency of delivery. Recent studies have demonstrated that PTD-mediated protein delivery is mediated by endocytotic pathways such as lipid raft-dependant macropinocytosis (8).…”

mentioning

confidence: 99%

Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides

Dixon

Osman

Morris

et al. 2016

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

128

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Protein transduction domains (PTDs) are powerful nongenetic tools that allow intracellular delivery of conjugated cargoes to modify cell behavior. Their use in biomedicine has been hampered by inefficient delivery to nuclear and cytoplasmic targets. Here we overcame this deficiency by developing a series of novel fusion proteins that couple a membrane-docking peptide to heparan sulfate glycosaminoglycans (GAGs) with a PTD. We showed that this GET (GAG-binding enhanced transduction) system could deliver enzymes (Cre, neomycin phosphotransferase), transcription factors (NANOG, MYOD), antibodies, native proteins (cytochrome C), magnetic nanoparticles (MNPs), and nucleic acids [plasmid (p)DNA, modified (mod)RNA, and small inhibitory RNA] at efficiencies of up to two orders of magnitude higher than previously reported in cell types considered hard to transduce, such as mouse embryonic stem cells (mESCs), human ESCs (hESCs), and induced pluripotent stem cells (hiPSCs). This technology represents an efficient strategy for controlling cell labeling and directing cell fate or behavior that has broad applicability for basic research, disease modeling, and clinical application.

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mentioning

confidence: 99%

Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides

Dixon

Osman

Morris

et al. 2016

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

128

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“…124 Although early results that were potentially affected by methodological artifacts suggested an energy-independent mode of uptake, it is now generally accepted that PTDs are primarily taken up by endocytosis. 121,125 The positive charge of the HIV1-TAT-PTD and other argininerich PTDs may help to concentrate the peptide on the cell surface by electrostatic interactions with negatively charged glycoproteins, 124,[126][127][128] similar to the mechanism described for transfection. Heparan sulfate proteoglycans (HSPGs) were proposed to assist in promoting attachment of HIV1-TAT protein and HIV1-TAT-PTD to the cell surface.…”

Section: ©2 0 1 1 L a N D E S B I O S C I E N C E D O N O T D I S Tmentioning

confidence: 99%

“…Internalization has consequently been suggested to occur via different routes of endocytosis, potentially including macropinocytosis, clathrin-and caveolin mediated endocytosis, as well as via a mode of entry by direct membrane penetration at high peptide concentrations. 125,[133][134][135] Although direct membrane penetration was proposed as a potential route of entry for PTDs alone, uptake of arginine-rich PTDs linked to macromolecular cargoes appears to occur solely by endocytosis. 127,134,136 Since routes of entry appear to be similar for most PTDs and delivery efficiency is subject to the same limitations, the potential applications of well-characterized PTDs are worth considering, rather than searching for the "best PTD."…”

Section: ©2 0 1 1 L a N D E S B I O S C I E N C E D O N O T D I S Tmentioning

confidence: 99%

Targeting antibodies to the cytoplasm

Marschall

Frenzel

Schirrmann

et al. 2011

mAbs

110

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“…Based on the high arginine content in the Tat sequence, Futaki et al synthesized a polypeptide composed solely of arginine residues [8], which can deliver macromolecules as efficiently as a Tat peptide [9,10]. Stearylated octaarginine (STR-R8) is a multifunctional device.…”

Section: Introductionmentioning

confidence: 99%

Topology of octaarginines (R8) or IRQ ligand on liposomes affects the contribution of macropinocytosis- and caveolae-mediated cellular uptake

Mudhakir¹,

Akita²,

Harashima³

2011

Reactive and Functional Polymers

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It was recently reported that liposomes modified with octaarginine (R8) and its analogue peptide (IRQRRRR: IRQ) are taken into NIH3T3 cells by unique pathways, macropinocytosis and caveolae-mediated endocytosis, respectively. This study evaluated the topology of these peptides as it relates to the uptake routes of liposomes, where they are modified either directly on the surface, or on the edge of a polyethylene glycol (PEG) spacer. The uptake mechanism of peptide-modified liposomes and peptide-modified PEG-liposomes was investigated by confocal laser scanning microscopy. To determine the contribution of clathrin-mediated endocytosis, macropinocytosis and caveolar endocytosis to the uptake of liposomes, the uptake was evaluated in the presence of these specific inhibitors. The uptake pathway changed from macropinocytosis to clathrin-mediated endocytosis when R8 was modified on the edge of a PEG spacer, indicating that the flexible display of R8 impaired the induction of macropinocytosis. However, the contribution of caveolae-mediated endocytosis increased when IRQ was conjugated to the distal end of the PEG chain, suggesting that flexible surface display enhanced IRQ recognition by the specific molecules in the caveolae. The present results demonstrate that topology control of the ligand affects the contribution of the entry pathway, depending on the uptake mechanism.

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Methodological and cellular aspects that govern the internalization mechanisms of arginine-rich cell-penetrating peptides

Cited by 338 publications

References 86 publications

Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides

Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides

Targeting antibodies to the cytoplasm

Topology of octaarginines (R8) or IRQ ligand on liposomes affects the contribution of macropinocytosis- and caveolae-mediated cellular uptake

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