TY - JOUR
T1 - Fast membrane association is a crucial factor in the peptide pep-1 translocation mechanism: a kinetic study followed by surface plasmon resonance.
AU - Henriques, Sonia T
AU - Castanho, Miguel A R B
AU - Pattenden, Leonard Keith
AU - Aguilar, Marie Isabel
PY - 2010
Y1 - 2010
N2 - The use of peptide carriers, termed cell-penetrating peptides or CPPs, has attracted much attention due to their potential for cellular delivery of hydrophilic molecules with pharmacological interest, overcoming the membrane barrier. These peptides are able to deliver attached cargos in a non-toxic manner, with the uptake mechanisms being either endosomally- or physically-driven. Pep-1 is a CPP of particular interest, not only due to outstanding delivery rates but also because its mechanism of membrane translocation is exclusively physically-driven which appears to be dependent on a very high affinity for the phospholipid bilayer in the cell membrane. In this report, pep-1-lipid interactions were further explored by characterization of the pep-1-lipid association/dissociation by surface plasmon resonance. Whereas a high affinity of pep-1 for lipid bilayers was observed in all conditions tested, negatively-charged phospholipids resulted in a larger peptide/lipid ratio. We also show that pep-1-membrane interaction is a fast process described by a multi-step model initiated by peptide adsorption, primarily governed by electrostatic attractions, and followed by peptide insertion in the hydrophobic membrane core. In the context of a cell-based process, the translocation of pep-1 is a physical mechanism promoted by peptide primary amphipathicity and asymmetric properties of the membrane. This explains the high efficiency rates of pep-1 when compared with other CPPs.
AB - The use of peptide carriers, termed cell-penetrating peptides or CPPs, has attracted much attention due to their potential for cellular delivery of hydrophilic molecules with pharmacological interest, overcoming the membrane barrier. These peptides are able to deliver attached cargos in a non-toxic manner, with the uptake mechanisms being either endosomally- or physically-driven. Pep-1 is a CPP of particular interest, not only due to outstanding delivery rates but also because its mechanism of membrane translocation is exclusively physically-driven which appears to be dependent on a very high affinity for the phospholipid bilayer in the cell membrane. In this report, pep-1-lipid interactions were further explored by characterization of the pep-1-lipid association/dissociation by surface plasmon resonance. Whereas a high affinity of pep-1 for lipid bilayers was observed in all conditions tested, negatively-charged phospholipids resulted in a larger peptide/lipid ratio. We also show that pep-1-membrane interaction is a fast process described by a multi-step model initiated by peptide adsorption, primarily governed by electrostatic attractions, and followed by peptide insertion in the hydrophobic membrane core. In the context of a cell-based process, the translocation of pep-1 is a physical mechanism promoted by peptide primary amphipathicity and asymmetric properties of the membrane. This explains the high efficiency rates of pep-1 when compared with other CPPs.
UR - http://onlinelibrary.wiley.com/doi/10.1002/bip.21367/pdf
U2 - 10.1002/bip.21367
DO - 10.1002/bip.21367
M3 - Article
SN - 0006-3525
VL - 94
SP - 314
EP - 322
JO - Biopolymers (Peptide Science)
JF - Biopolymers (Peptide Science)
IS - 3
ER -