title: Single-molecule imaging of the association of the cell-penetrating peptide Pep-1 to model membranes.
abstract: Pep-1 is an amphiphatic peptide that can form noncovalent complexes with a cargo protein with subsequent delivery into a live cell. In this study, the behavior of Pep-1 was directly visualized by fluorescent imaging techniques at the single-molecule level of sensitivity. The interactions of Pep-1 and two of its labeled fluorescent analogues with large and cell-sized giant unilamellar vesicles and supported bilayers are reported. The role of the bilayer charge and ionic strength of the medium were examined. Pep-1 caused fusion and association of vesicles, and it perturbed the vesicle's membrane. The association of the peptide with neutral bilayers was promoted by anchoring of the cysteamine moiety. The association of the peptide with the structural defects of the neutral membrane was very efficient. The electrostatic forces were shown to be important for the association of the peptide only in low ionic strength solutions and were completely diminished at physiological ionic strength. Pep-1 did not induce the association to the model membrane of a number of proteins chosen to exhibit a range of properties. The results suggest that Pep-1 assisted delivery of cargo in living cells may result from cooperative effects.
authors.affiliation: Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6323, USA.
authors.forename: Alexey
authors.initials: A
authors.lastname: Sharonov
authors.forename: Robin M
authors.initials: RM
authors.lastname: Hochstrasser
mesh_terms: D000595:Amino Acid Sequence; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008567:Membranes, Artificial; C503924:Pep-1 peptide; D010455:Peptides; D003543:Cysteamine