Kinetic process of beta-amyloid formation via membrane binding.
Recently we have studied thermodynamics of membrane-mediated beta-amyloid formation in equilibrium experiments using penetratin-lipid mixtures. The results showed that penetratin bound to the membrane interface in the alpha-helical conformation when the peptide/lipid (P/L) ratios were below a lipid-dependent critical value P/L*. When P/L reached P/L*, small beta-aggregates emerged, which served as the nuclei for large beta-aggregates. Here we studied the corresponding kinetic process to understand the potential barriers for the membrane-mediated beta-amyloid formation. We performed kinetic experiments using giant unilamellar vesicles made of 7:3 DOPC/DOPG. The observed time behavior of individual giant unilamellar vesicles, although complex, exhibited the physical effects seen in equilibrium experiments. Most interestingly, a potential barrier appeared to block penetratin from translocating across the bilayer. As a result, the kinetic value for the critical threshold P/L* is roughly one-half of the value measured in equilibrium where peptides bind symmetrically on both sides of lipid bilayers. We also investigated the similarity and differences between the charged and neutral lipids in their interactions with penetratin. We reached an important conclusion that the bound states of peptides in lipid bilayers are largely independent of the charge on the lipid headgroups.
Sun
Yen
Y
Department of Physics and Astronomy, Rice University, Houston, Texas, USA.
Lee
Chang-Chun
CC
Chen
Tzu-Hsuan
TH
Huang
Huey W
HW
D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D002942:Circular Dichroism; D007700:Kinetics; D008856:Microscopy, Fluorescence; D010713:Phosphatidylcholines / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D055672:Static Electricity; D053835:Unilamellar Liposomes / Q000378:metabolism; D014961:X-Ray Diffraction
D016229:Amyloid beta-Peptides; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D053835:Unilamellar Liposomes; C051388:1,2-dioleoyl-sn-glycero-3-phosphoglycerol; C017251:1,2-oleoylphosphatidylcholine