Translocation of positively charged copoly(Lys/Tyr) across phospholipid membranes.
Much attention has recently been paid to the study of positively charged polypeptides as a possible carrier for therapeutic protein or DNA delivery to cells. In this study, we have investigated the translocation of positively charged copoly(Lys/Tyr) (MW=72000, DP=385) across lipid membranes constituted from egg-phosphatidylcholine (EPC), dioleoyl-phosphatidylethanolamine (DOPE), as well as soybean phospholipids (SBPL) using zeta potential method, circular dichroism spectroscopy (CD), electrophysiology technique, fluorescence spectroscopy, and confocal laser scanning microscopy. Results of zeta potentials show that copoly(Lys/Tyr) associate with lipid membranes and become gradually saturated on the membranes either hydrophobically or electrostatically or both. CD studies demonstrate that the copoly(Lys/Tyr) takes and remains beta-sheet conformation during its interaction with liposome membranes, indicating that the translocation process should be carpet-mode like. Data from the electrophysiology technique reveal that positively charged copoly(Lys/Tyr) can cause transmembrane currents under an applied voltage, confirming its transfer across lipid membranes. Fluorescence spectroscopy results display a three-step mechanism of translocation across membrane: adsorption, transportation, and desorption, which has been verified by results from confocal laser scanning microscopy. We provided the first direct observation that the positively charged polypeptides, copoly(Lys/Tyr), can translocate through SBPL and EPC/DOPE lipid bilayer membranes. In addition, we found that the translocation efficiency of copoly(Lys/Tyr) was higher on the EPC/DOPE lipid membrane than on the SBPL lipid membrane.
Liu
Shaoqian
S
College of Chemistry and chemical Engineering, Central South University, Changsha 410083, China. liushao2001@yahoo.com.cn
Shibata
Akira
A
Ueno
Satoru
S
Huang
Ying
Y
Wang
Yifan
Y
Li
Yuanjian
Y
D004574:Electromagnetic Fields; D018274:Electroporation / Q000379:methods*; D008051:Lipid Bilayers / Q000737:chemistry* / Q000528:radiation effects; D008081:Liposomes / Q000737:chemistry* / Q000528:radiation effects; D008560:Membrane Fluidity / Q000528:radiation effects*; D008563:Membrane Lipids / Q000737:chemistry* / Q000528:radiation effects; D008567:Membranes, Artificial; D009038:Motion; D010455:Peptides / Q000737:chemistry* / Q000528:radiation effects; D010743:Phospholipids / Q000737:chemistry* / Q000528:radiation effects; D011108:Polymers; D055672:Static Electricity
D008051:Lipid Bilayers; D008081:Liposomes; D008563:Membrane Lipids; D008567:Membranes, Artificial; D010455:Peptides; D010743:Phospholipids; D011108:Polymers; C028311:poly-L-lysyltyrosine