scholarly journals Free Energy of Translocating an Arginine-Rich Cell-Penetrating Peptide across a Lipid Bilayer Suggests Pore Formation

2013 ◽  
Vol 104 (2) ◽  
pp. 412-420 ◽  
Author(s):  
Kun Huang ◽  
Angel E. García
2005 ◽  
Vol 89 (6) ◽  
pp. 4300-4309 ◽  
Author(s):  
Thomas Plénat ◽  
Sylvie Boichot ◽  
Patrice Dosset ◽  
Pierre-Emmanuel Milhiet ◽  
Christian Le Grimellec

2011 ◽  
Vol 112 (1) ◽  
pp. 178-183 ◽  
Author(s):  
Shuhei Kawamoto ◽  
Takeshi Miyakawa ◽  
Masako Takasu ◽  
Ryota Morikawa ◽  
Tatsuki Oda ◽  
...  

Author(s):  
Evgeniya Trofimenko ◽  
Gianvito Grasso ◽  
Mathieu Heulot ◽  
Nadja Chevalier ◽  
Marco A. Deriu ◽  
...  

SummaryCell-penetrating peptides (CPPs) allow intracellular delivery of cargo molecules. CPPs provide efficient methodology to transfer bioactive molecules in cells, in particular in conditions when transcription or translation of cargo-encoding sequences is not desirable or achievable. The mechanisms allowing CPPs to enter cells are ill-defined and controversial. This work identifies potassium channels as key regulators of cationic CPP translocation. Using a CRISPR/Cas9-based screening, we discovered that KCNQ5, KCNN4, and KCNK5 positively modulate CPP cellular direct translocation by reducing transmembrane potential (Vm). Cationic CPPs further decrease the Vm to megapolarization values (about −150 mV) leading to the formation of ∼2 nm-wide water pores used by CPPs to access the cell’s cytoplasm. Pharmacological manipulation to lower transmembrane potential boosted CPPs cellular uptake in zebrafish and mouse models. Besides identifying the first genes that regulate CPP translocation, this work characterizes key mechanistic steps used by CPPs to cross cellular membrane. This opens the ground for pharmacological strategies augmenting the susceptibility of cells to capture CPP-linked cargos in vitro and in vivo.


2021 ◽  
Vol 203 (9) ◽  
Author(s):  
Farzana Hossain ◽  
Hideo Dohra ◽  
Masahito Yamazaki

ABSTRACT An antimicrobial peptide (AMP) derived from lactoferricin B, LfcinB(4–9) (RRWQWR), and lissamine rhodamine B red-labeled peptide [Rh-LfcinB(4–9)] exhibit strong antimicrobial activities, and they can enter Escherichia coli cells without damaging the cell membranes. Thus, these peptides are cell-penetrating peptide (CPP)-type AMPs. In this study, to elucidate the effect of the membrane potential (Δφ) on the action of the CPP-type AMP Rh-LfcinB(4–9), we investigated the interactions of Rh-LfcinB(4–9) with single E. coli cells and spheroplasts containing calcein in the cytosol using confocal laser scanning microscopy. At low peptide concentrations, Rh-LfcinB(4–9) entered the cytosol of single E. coli cells and spheroplasts without damaging the cell membranes, and the H+ ionophore carbonyl cyanide m-chlorophenylhydrazone (CCCP) suppressed its entry. Studies using the time-kill method indicate that these low concentrations of peptide exhibit antimicrobial activity, but CCCP inhibits this activity. Next, we investigated the effect of Δφ on the interaction of Rh-LfcinB(4–9) with single giant unilamellar vesicles (GUVs) comprising E. coli polar lipid extracts and containing a fluorescent probe, Alexa Fluor 647 hydrazide. At low concentrations (0.2 to 0.5 μM), Rh-LfcinB(4–9) showed significant entry into the single GUV lumen without pore formation in the presence of Δφ. The fraction of entry of the peptide increased with increasing negative membrane potential, indicating that the rate of peptide entry into the GUV lumen increased with increasing negative membrane potential. These results indicate that Δφ enhances the entry of Rh-LfcinB(4–9) into single E. coli cells, spheroplasts, and GUVs and its antimicrobial activity. IMPORTANCE Bacterial cells have a membrane potential (Δφ), but the effect of Δφ on the action of cell-penetrating peptide-type antimicrobial peptides (AMPs) is not clear. Here, we investigated the effect of Δφ on the action of a fluorescent probe-labeled AMP derived from lactoferricin B, Rh-LfcinB(4–9). At low peptide concentrations, Rh-LfcinB(4–9) enters the cytosol of Escherichia coli cells and spheroplasts without damaging their cell membrane, but a protonophore suppresses this entry and its antimicrobial activity. The rate of entry of Rh-LfcinB(4–9) into the giant unilamellar vesicles (GUVs) comprising E. coli lipids without pore formation increases with increasing Δφ. These results indicate that Δφ enhances the antimicrobial activity of Rh-LfcinB(4–9) and, hence, LfcinB(4–9) by increasing the rate of their entry into the cytosol.


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