Synthetic copoly(Lys/Phe) and poly(Lys) translocate through lipid bilayer membranes.
Several membrane-transporting peptides (MTP) containing basic amino acid residues such as Lys and Arg that carry macromolecules such as DNA and proteins across cell plasma membranes by an unknown mechanism have been actively studied. On the basis of these results, we have been investigating the translocation ability of synthetic polypeptides, copoly(Lys/Phe) and poly(Lys), through negatively charged phospholipid (soybean phospholipid (SBPL)) bilayer membranes by zeta potential analysis, circular dichroism (CD) spectroscopy, fluorescence spectroscopy, an electrophysiology technique, and confocal laser scanning microscopy (CLSM). The binding of these polypeptides to the membrane, which is the first step for translocation across the membrane, resulted in the conformational transition of the polypeptide from a random coil form or helix-poor form to a helix-rich form. The fluorescence studies demonstrated that the time-dependent decrease in the fluorescence intensities of the FITC-labeled polypeptides bound to the SBPL liposome reflected translocation of the polypeptide across the lipid bilayer with the low dielectric constant. Both the rate constant and the efficiency of the polypeptide translocation across the lipid bilayer were greater for copoly(Lys/Phe) than for poly(Lys). These results suggest that the random incorporation of the hydrophobic Phe residue into the positively charged Lys chain results in a lowering of the potential barrier for passage of the polypeptide in the hydrophobic core portion of the lipid bilayer. We presented the first direct observation that the positively charged polypeptides, copoly(Lys/Phe) (MW: 41,500) and poly(Lys) (MW: 23,400), could translocate across the lipid bilayer membrane.
Shibata
Akira
A
Faculty of Pharmaceutical Sciences, Tokushima University, Shomachi, Tokushima 770-8505, Japan. ashibata@ph.tokushima-u.ac.jp
Murata
Syuichi
S
Ueno
Satoru
S
Liu
Shaoqian
S
Futaki
Shiroh
S
Baba
Yoshinobu
Y
D000327:Adsorption; D001692:Biological Transport; D002942:Circular Dichroism; D008051:Lipid Bilayers / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry; D018613:Microscopy, Confocal; D010455:Peptides / Q000378:metabolism*; D011107:Polylysine / Q000378:metabolism*; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence
D008051:Lipid Bilayers; D008081:Liposomes; D010455:Peptides; D011107:Polylysine