Cell‐penetrating peptides recruit type A scavenger receptors to the plasma membrane for cellular delivery of nucleic acids

2016 ◽  
Vol 31 (3) ◽  
pp. 975-988 ◽  
Author(s):  
Carmen Juks ◽  
Annely Lorents ◽  
Piret Arukuusk ◽  
Ülo Langel ◽  
Margus Pooga
Keyword(s):  
Plasma Membrane ◽  
Nucleic Acids ◽  
Type A ◽  
Cell Penetrating Peptides ◽  
Scavenger Receptors ◽  
Cellular Delivery ◽  
Cell Penetrating

scholarly journals Low concentration thresholds of plasma membranes for rapid energy-independent translocation of a cell-penetrating peptide

2009 ◽  
Vol 420 (2) ◽  
pp. 179-191 ◽  
Author(s):  
Catherine L. Watkins ◽  
Dirk Schmaljohann ◽  
Shiroh Futaki ◽  
Arwyn T. Jones
Keyword(s):  
Plasma Membrane ◽  
Cell Penetrating Peptides ◽  
Cell Penetrating Peptide ◽  
Cholesterol Depletion ◽  
Full Potential ◽  
Adherent Cells ◽  
High Porosity ◽  
Cellular Delivery ◽  
Suspension Cells ◽  
Cell Penetrating

The exact mechanisms by which cell-penetrating peptides such as oligo-arginines and penetratin cross biological membranes has yet to be elucidated, but this is required if they are to reach their full potential as cellular delivery vectors. In the present study, qualitative and quantitative analysis of the influence of temperature, peptide concentration and plasma membrane cholesterol on the uptake and subcellular distribution of the model cell-penetrating peptide octa-arginine was performed in a number of suspension and adherent cell lines. When experiments were performed on ice, the peptide at 2 μM extracellular concentration efficiently entered and uniformly labelled the cytoplasm of all the suspension cells studied, but a 10-fold higher concentration was required to observe similar results in adherent cells. At 37 °C and at higher peptide concentrations, time-lapse microscopy experiments showed that the peptide rapidly penetrated the entire plasma membrane of suspension cells, with no evidence of a requirement for nucleation zones to promote this effect. Cholesterol depletion with methyl-β-cyclodextrin enhanced translocation of octa-arginine across the plasma membrane of suspension cells at 37 °C, but decreased overall peptide accumulation. Under the same conditions in adherent cells this agent had no effect on peptide uptake or distribution. Cholesterol depletion increased the overall accumulation of the peptide at 4 °C in KG1a cells, but this effect could be reversed by re-addition of cholesterol as methyl-β-cyclodextrin–cholesterol complexes. The results highlight the relatively high porosity of the plasma membrane of suspension cells to this peptide, especially at low temperatures, suggesting that this feature could be exploited for delivering bioactive entities.

Addressing the challenges of cellular delivery and bioavailability of peptide nucleic acids (PNA)

2005 ◽  
Vol 38 (4) ◽  
pp. 345-350 ◽  
Author(s):  
Peter E. Nielsen
Keyword(s):  
Nucleic Acids ◽  
Peptide Nucleic Acid ◽  
Peptide Nucleic Acids ◽  
Cell Penetrating Peptides ◽  
Endosomal Escape ◽  
Cellular Delivery ◽  
Peptide Conjugation ◽  
Cell Penetrating ◽  
Antisense Peptide

1. Introduction 3452. Peptide nucleic acid (PNA) 3463. ‘Cell penetrating peptides’ (CPPs) 3464. Endosomal escape 3475. Cellular delivery of PNA 3476.In vivobioavailability of PNA 3497. References 350Recent results on the cellular delivery of antisense peptide nucleic acids (PNA) via peptide conjugation is briefly discussed, in particular in the context of endosomal entrapment and escape.

scholarly journals Temperature-, concentration- and cholesterol-dependent translocation of L- and D-octa-arginine across the plasma and nuclear membrane of CD34+ leukaemia cells

2007 ◽  
Vol 403 (2) ◽  
pp. 335-342 ◽  
Author(s):  
Marjan M. Fretz ◽  
Neal A. Penning ◽  
Saly Al-Taei ◽  
Shiroh Futaki ◽  
Toshihide Takeuchi ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Penetrating Peptides ◽  
Threshold Concentration ◽  
Membrane Cholesterol ◽  
Cellular Delivery ◽  
Uptake Mechanism ◽  
Peptide Concentration ◽  
Cell Penetrating ◽  
Plasma Membrane Cholesterol ◽  
In Cells

Delineating the mechanisms by which cell-penetrating peptides, such as HIV-Tat peptide, oligoarginines and penetratin, gain access to cells has recently received intense scrutiny. Heightened interest in these entities stems from their ability to enhance cellular delivery of associated macromolecules, such as genes and proteins, suggesting that they may have widespread applications as drug-delivery vectors. Proposed uptake mechanisms include energy-independent plasma membrane translocation and energy-dependent vesicular uptake and internalization through endocytic pathways. In the present study, we investigated the effects of temperature, peptide concentration and plasma membrane cholesterol levels on the uptake of a model cell-penetrating peptide, L-octa-arginine (L-R8) and its D-enantiomer (D-R8) in CD34+ leukaemia cells. We found that, at 4–12 °C, L-R8 uniformly labels the cytoplasm and nucleus, but in cells incubated with D-R8 there is additional labelling of the nucleolus which is still prominent at 30 °C incubations. At temperatures between 12 and 30 °C, the peptides are also localized to endocytic vesicles which consequently appear as the only labelled structures in cells incubated at 37 °C. Small increases in the extracellular peptide concentration in 37 °C incubations result in a dramatic increase in the fraction of the peptide that is localized to the cytosol and promoted the binding of D-R8 to the nucleolus. Enhanced labelling of the cytosol, nucleus and nucleolus was also achieved by extraction of plasma membrane cholesterol with methyl-β-cyclodextrin. The data argue for two, temperature-dependent, uptake mechanism for these peptides and for the existence of a threshold concentration for endocytic uptake that when exceeded promotes direct translocation across the plasma membrane.

Peptide-based delivery of nucleic acids: design, mechanism of uptake and applications to splice-correcting oligonucleotides

2007 ◽  
Vol 35 (1) ◽  
pp. 53-55 ◽  
Author(s):  
S. Abes ◽  
H. Moulton ◽  
J. Turner ◽  
P. Clair ◽  
J.P. Richard ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cell Membrane ◽  
Cell Surface ◽  
Nucleic Acids ◽  
Peptide Nucleic Acids ◽  
Basic Amino Acid ◽  
Cell Penetrating Peptides ◽  
Cellular Delivery ◽  
Cell Penetrating ◽  
Design Mechanism

CPPs (cell-penetrating peptides) have given rise to much interest for the delivery of biomolecules such as peptides, proteins or ONs (oligonucleotides). CPPs and their conjugates were initially thought to translocate through the cell membrane by a non-endocytotic mechanism which has recently been re-evaluated. Basic-amino-acid-rich CPPs first interact with cell-surface proteoglycans before being internalized by endocytosis. Sequestration and degradation in endocytotic vesicles severely limits the cytoplasmic and nuclear delivery of the conjugated biomolecules. Accordingly, splicing correction by CPP-conjugated steric-block ON analogues is inefficient in the absence of endosomolytic agents. New arginine-rich CPPs allowing efficient splicing correction by conjugated PNAs (peptide nucleic acids) or PMO (phosphorodiamidate morpholino oligomer) steric blockers in the absence of endosomolytic agents have recently been defined in our group and are currently being characterized. They offer promising leads for the development of efficient cellular delivery vectors for therapeutic steric-block ON analogues.

Cell-penetrating peptides for the delivery of nucleic acids

2012 ◽  
Vol 9 (7) ◽  
pp. 823-836 ◽  
Author(s):  
Taavi Lehto ◽  
Kaido Kurrikoff ◽  
Ülo Langel
Keyword(s):  
Nucleic Acids ◽  
Cell Penetrating Peptides ◽  
Cell Penetrating

Translocation of cell-penetrating peptides across the plasma membrane is controlled by cholesterol and microenvironment created by membranous proteins

2014 ◽  
Vol 192 ◽  
pp. 103-113 ◽  
Author(s):  
Janely Pae ◽  
Pille Säälik ◽  
Laura Liivamägi ◽  
Dmitri Lubenets ◽  
Piret Arukuusk ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Penetrating Peptides ◽  
Cell Penetrating

Cell Penetrating Mechanism of Bax Inhibiting Peptides.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4298-4298
Author(s):  
Jose A. Gomez ◽  
Tomoyuki Yoshida ◽  
Minh Lam ◽  
Clark W. Distelhorst ◽  
Shigemi Matsuyama
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Fluorescent Dyes ◽  
Cultured Cells ◽  
Cell Penetrating Peptides ◽  
Dependent Manner ◽  
Cell Penetration ◽  
New Members ◽  
Cell Penetrating ◽  
High Degree

Abstract Plasma membrane is known to have a high degree of selectivity for molecular trafficking, and it does not allow the penetration of peptides larger than 3 amino acids. Previously known exceptions of large peptides that penetrate the plasma membrane are the Arginine rich peptides such as human immunodeficiency virus (HIV)-tat peptides. However, the mechanism of cell penetration of these peptides is largely unknown. Bax Inhibiting Peptides (BIP) are penta-peptides derived from the Bax binding domain of Ku70. At present, three types of BIP have been developed. Those are: VPMLK, VPTLK, and VPALR. All of these three BIPs directly bind Bax and inhibit Bax-mediated cell death in cultured cells as well as in animal study. Surprisingly, BIPs are cell permeable and autonomously enter the cytoplasm of the cells within 1 hr. Therefore BIPs are recognized as new members of cell penetration peptides. In this study, we investigated the mechanism of cell penetration of BIPs. DAMI cells (a human megakaryocyte cell line) and HeLa cells were used to investigate the detailed mechanism of cell penetration of BIPs. To detect the cell entry of BIPs, fluorescent dyes (fluorescein or tetramethylrodhamine) were conjugated to the N-terminus of BIPs and the cytoplasmic localization of BIPs was confirmed by confocal microscopy. Cell Penetration activities of BIPs were detected at 1 uM concentration in the culture medium. The significant accumulation of BIPs in the cytoplasm were detected within 1 hour of incubation both at 4 °C and 37 °C, suggesting that ATP-independent mechanism of cell penetration of BIP exists. However, cellular uptake of BIPs reaches plateau at 100 uM at 4 °C, whereas it increases in a dose dependent manner up to 1 mM at 37 °C without any sign of cytotoxicity. These results suggest that there are at least two mechanisms contributing to the cell penetration of BIPs that are, “ATP-independent (4 °C)” and “ATP-dependent (37 °C)” mechanisms. In addition to BIPs, we generated a series of mutated BIPs that do not bind Bax but retain cell-penetrating activities. We performed competition assay using fluorescence dye-labeled and non-labeled BIP (and the mutant BIPs), and the preliminary results suggest that there is a specific receptor for each peptide for its delivery into the cells. Our data also indicates that BIPs can deliver a cargo molecule (e.g. fluorescent dye) with at least the same molecular weight. Unlike other cell penetrating peptides, BIP has minimum toxicity due to its nature to inhibit Bax-mediated cell death. Along with the new data showing that BIP protects cells from pathological damages in cell culture and animal model, we will discuss the potential application of BIPs as a new type of drug delivery tool.

scholarly journals Role of Cell-Penetrating Peptides in Intracellular Delivery of Peptide Nucleic Acids Targeting Hepadnaviral Replication

2017 ◽  
Vol 9 ◽  
pp. 162-169 ◽  
Author(s):  
Bénédicte Ndeboko ◽  
Narayan Ramamurthy ◽  
Guy Joseph Lemamy ◽  
Catherine Jamard ◽  
Peter E. Nielsen ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Peptide Nucleic Acids ◽  
Intracellular Delivery ◽  
Cell Penetrating Peptides ◽  
Cell Penetrating
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