Low concentration thresholds of plasma membranes for rapid energy-independent translocation of a cell penetrating peptide - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Biochemical Journal Année : 2009

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

Catherine L. Watkins
  • Fonction : Auteur
Dirk Schmaljohann
  • Fonction : Auteur
Shiroh Futaki
Arwyn T. Jones
  • Fonction : Auteur correspondant
  • PersonId : 870043

Connectez-vous pour contacter l'auteur

Résumé

The exact mechanisms by which cell penetrating peptides such as oligoarginines 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. Here, 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 octaarginine 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 studied suspension cells but a ten-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 translocated 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 octaarginine 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 this feature could be exploited for delivering bioactive entities.

Mots clés

Fichier principal
Vignette du fichier
PEER_stage2_10.1042%2FBJ20090042.pdf (5.48 Mo) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)
Loading...

Dates et versions

hal-00479146 , version 1 (30-04-2010)

Identifiants

Citer

Catherine L. Watkins, Dirk Schmaljohann, Shiroh Futaki, Arwyn T. Jones. Low concentration thresholds of plasma membranes for rapid energy-independent translocation of a cell penetrating peptide. Biochemical Journal, 2009, 420 (2), pp.179-189. ⟨10.1042/BJ20090042⟩. ⟨hal-00479146⟩

Collections

PEER
32 Consultations
156 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More