The Efficacies of Cell‐Penetrating Peptides in Accumulating in Large Unilamellar Vesicles Depend on their Ability To Form Inverted Micelles

Swiecicki, Jean-Marie; Bartsch, Annika; Tailhades, Julien; Pisa, Margherita Di; Heller, Benjamin J.; Chassaing, Gérard; Mansuy, Christelle; Burlina, Fabienne; Lavielle, Solange

doi:10.1002/cbic.201300742

Cited by 55 publications

(100 citation statements)

References 66 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…7), anionic oxidized lipids, including anionic oxidized phosphatidylcholine species, directly recruit cationic CPPs to the cell surface and mediate the transfer of the peptide across the bilayer by formation of structures such as inverted micelles. This model is consistent with previously proposed translocation models involving inverted micelles composed of other anionic species (23,26,44,65,66). An important distinction, however, is that membrane oxidation is a key variable that can explain how anionic lipids are presented on the cell surface.…”

Section: Discussionsupporting

confidence: 92%

Membrane Oxidation Enables the Cytosolic Entry of Polyarginine Cell-penetrating Peptides

Wang

Sun

Muthukrishnan

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

Arginine-rich peptides can penetrate cells and consequently be used as delivery agents in various cellular applications. The activity of these reagents is often context-dependent, and the parameters that impact cell entry are not fully understood, giving rise to variability and limiting progress toward their usage. Herein, we report that the cytosolic penetration of linear polyarginine peptides is dependent on the oxidation state of the cell. In particular, we find that hypoxia and cellular antioxidants inhibit cell penetration. In contrast, oxidants promote cytosolic cell entry with an efficiency proportional to the level of reactive oxygen species generated within membranes. Moreover, an antibody that binds to oxidized lipids inhibits cell penetration, whereas extracellularly administered pure oxidized lipids enhance peptide transport into cells. Overall, these data indicate that oxidized lipids are capable of mediating the transport of polyarginine peptides across membranes. These data may also explain variability in cell-penetrating peptide performance in different experimental conditions. These new findings therefore provide new opportunities for the rational design of future cell-permeable compounds and for the optimization of delivery protocols.

show abstract

Section: Discussionsupporting

confidence: 92%

Membrane Oxidation Enables the Cytosolic Entry of Polyarginine Cell-penetrating Peptides

Wang

Sun

Muthukrishnan

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…As indicated (Figure 6), the fast or peptide penetration kinetic phase can be fit by a single-exponential function with a time constant of 10.5 ± 2.2 minutes. This value is similar to those reported in the literature for TAT [40,46,47], providing further supporting evidence for our assignment.…”

Section: Resultssupporting

confidence: 92%

“…Here we show, using TAT as an example, that this concern can be alleviated by using Phe CN fluorescence as a probe to follow the kinetics of CPP membrane penetration. Specifically, we replaced the N-terminal Tyr residue of TAT with Phe CN , (the resultant peptide is hereinafter referred to as F CN -TAT), which is expected to cause only a minimum perturbation to the peptide as Tyr and Phe CN are similar in size, and we exploited the pH-dependence of the Phe CN fluorescence to quantitatively assess the penetration kinetics of TAT across a model membrane [40]. Interestingly, we find that under our experimental conditions TAT first translocates across the membrane on a timescale of minutes and then causes membrane leakage on a timescale of hours.…”

Section: Introductionmentioning

confidence: 99%

Sensing pH via p-cyanophenylalanine fluorescence: Application to determine peptide pKa and membrane penetration kinetics

Pazos

Ahmed

Berríos

et al. 2015

Analytical Biochemistry

View full text Add to dashboard Cite

We expand the spectroscopic utility of a well-known infrared and fluorescence probe, p-cyanophenylalanine, by showing that it can also serve as a pH sensor. This new application is based on the notion that the fluorescence quantum yield of this unnatural amino acid, when placed at or near the N-terminal end of a polypeptide, depends on the protonation status of the N-terminal amino group of the peptide. Using this pH sensor, we are able to determine the N-terminal pKa values of nine tripeptides and also the membrane penetration kinetics of a cell-penetrating peptide. Taken together, these examples demonstrate the applicability of using this unnatural amino acid fluorophore to study pH-dependent biological processes or events that accompany a pH change.

show abstract

“…Yet, more recently GVs derived from the plasma membrane (lacking endocytosis machinery) were used by Pooga and collaborators that demonstrate the ability of uptake of 6 CPPs, including penetratin. 9 In more recent years, translocation of penetratin, polyarginines and other cationic peptides was demonstrated in LUVs using uorescence studies 14,15 and mass spectrometry approaches 16 further corroborating the idea that CPPs can directly translocate through pure lipid membranes.…”

Section: Introductionmentioning

confidence: 89%

Label-free quantification of cell-penetrating peptide translocation into liposomes

Jobin

Alves

2016

Anal. Methods

View full text Add to dashboard Cite

Cell penetrating peptides (CPPs) are small molecules capable of crossing lipid membranes and transporting cargos of varied sizes and nature inside cells. Two cellular uptake mechanisms have been identified: endocytosis and direct translocation through the cellular membrane. Several methods have been proposed to follow and quantify CPP internalization ability both in cells and liposomes, most of them requiring the addition of some sort of label to the CPP. Herein we propose a protocol to test and quantify the translocation ability of CPPs through liposomes by measuring their intrinsic Trp fluorescence and their quenching by acrylamide. Although restrained to CPPs that possess at least one Trp residue in their sequence, the protocol remains applicable to a great number of CPPs. The protocol was applied to three CPPs, including one of the first discovered (penetratin), for their internalization into zwitterionic and anionic liposomes and upon different incubation times. While penetratin and one of its analogs were able to translocate into anionic liposomes, none of the peptides translocated into zwitterionic ones. The newly developed protocol allows for the detection of nanomolar amounts of CPPs inside liposomes. The results obtained are in very good agreement with those determined by other quantification methods.

show abstract

The Efficacies of Cell‐Penetrating Peptides in Accumulating in Large Unilamellar Vesicles Depend on their Ability To Form Inverted Micelles

Cited by 55 publications

References 66 publications

Membrane Oxidation Enables the Cytosolic Entry of Polyarginine Cell-penetrating Peptides

Membrane Oxidation Enables the Cytosolic Entry of Polyarginine Cell-penetrating Peptides

Sensing pH via p-cyanophenylalanine fluorescence: Application to determine peptide pKa and membrane penetration kinetics

Label-free quantification of cell-penetrating peptide translocation into liposomes

Contact Info

Product

Resources

About