Simultaneous removal of thiolated membrane proteins resulting in nanostructured lipid layers.
Self-organization of membrane-embedded peptides and proteins causes the formation of lipid mesostructures in the membranes. One example is purple membranes (PM), which consist of lipids and bacteriorhodopsin (BR) as the only protein component. The BRs form a hexagonal crystalline lattice. A complementary structure is formed by the lipids. Employing BR and PM as an example, we report a method where major parts of the mesoscopic self-assembled protein structures can be extracted from the lipid bilayer membrane. A complementary lipid nanostructure remains on the substrate. To remove such a large number of thiolated proteins simultaneously by applying a mechanical force, they are first reacted at physiological conditions with gold nanoparticles, and then a thin gold film is sputtered onto them that fuses with the gold nanoparticles forming a uniform layer, which finally can be lifted off. In this step, all of the previously gold-labeled proteins are pulled out of the membrane simultaneously. A stable lipid nanostructure is obtained on the mica substrate. Its stability is due to either binding of the lipids to the substrate through ionic bonds or to enough residual proteins to stabilize the lipid nanostructure against reorganization. This method may be applied easily and efficiently wherever thiolated proteins or peptides are employed as self-assembling and structure-inducing units in lipid membranes.
Wu
Aiguo
A
Faculty of Chemistry, University of Marburg, Hans-Meerwein-Strasse, D-35032 Marburg, Germany.
Jia
Zhihong
Z
Schaper
Andreas
A
Noll
Frank
F
Hampp
Norbert A
NA
D001436:Bacteriorhodopsins / Q000737:chemistry*; D006046:Gold / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D013438:Sulfhydryl Compounds / Q000737:chemistry*
D008051:Lipid Bilayers; D013438:Sulfhydryl Compounds; D001436:Bacteriorhodopsins; D006046:Gold