Consequences of nonlytic membrane perturbation to the translocation of the cell penetrating peptide ppep-1 in lipidic vesicles

Troeira Henriques, Sonia and Castanho, Miguel Augusto Rico Botas (2004) Consequences of nonlytic membrane perturbation to the translocation of the cell penetrating peptide ppep-1 in lipidic vesicles. Biochemistry, 43 30: 9716-9724. doi:10.1021/bi036325k


Author Troeira Henriques, Sonia
Castanho, Miguel Augusto Rico Botas
Title Consequences of nonlytic membrane perturbation to the translocation of the cell penetrating peptide ppep-1 in lipidic vesicles
Journal name Biochemistry   Check publisher's open access policy
ISSN 0006-2960
Publication date 2004-08-03
Sub-type Article (original research)
DOI 10.1021/bi036325k
Volume 43
Issue 30
Start page 9716
End page 9724
Total pages 19
Place of publication Washington, D.C., U.S.A.
Publisher American Chemical Society
Language eng
Subject 0601 Biochemistry and Cell Biology
Formatted abstract
The action of the cell penetrating pep-1 at the molecular level is not clearly understood. The ability of the peptide to induce (1) vesicle aggregation, (2) lipidic fusion, (3) anionic lipid segregation, (4) pore or other lytic structure formation, (5) asymmetric lipidic flip-flop, and (6) peptide translocation across the bilayers in large unilamellar vesicles was studied using photophysical methodologies mainly related to fluorescence spectroscopy. Neflometry and turbidimetry techniques show that clustering of vesicles occurs in the presence of the peptide in a concentration- and anionic lipid content-dependent manner. Results from Forstër resonance energy transfer-based methodologies prove lipidic fusion and anionic lipid segregation, but no evidence for pores or other lytic structures was found. Asymmetric lipid flip-flop was not detected either. A specific method related to the quenching of the rhodamine-labeled lipids by pep-1 was developed to study the eventual translocation of the peptide. Translocation does not occur in symmetrical neutral and negatively charged vesicles, except when a valinomycin-induced transmembrane potential exists. Our work strongly suggests that the main driving force for peptide translocation is charge asymmetry between the outer and inner leaflet of biological membranes and reveals that pep-1 is able to perturb membranes without being cytotoxic. This nonlytic perturbation is probably mandatory for translocation to occur.
Keyword Biological membranes
Cells
Cytotoxic
Peptide translocation
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
Institute for Molecular Bioscience - Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 66 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 65 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 24 Mar 2010, 00:50:39 EST by Ms May Balasaize on behalf of Institute for Molecular Bioscience