Design of supported membranes tethered via metal-affinity ligand-receptor pairs.
Model lipid layers are very promising in investigating the complex network of recognition, transport and signaling processes at membranes. We have developed a novel and generic approach to create supported lipid membranes tethered by metal-affinity binding. By self-assembly we have generated various interfaces that display histidine sequences (6xHis) via polymer spacers. These histidine-functionalized interfaces are designed to allow specific docking and fusion of vesicles containing metal-chelating lipids. By means of surface plasmon resonance and atomic force microscopy we analyzed the formation and subsequently the structure of these solid-supported membranes. Although the affinity constant of single ligand-receptor pairs is only in the micromolar range, very stable immobilization of these membranes was observed. This behavior can be explained by multivalent interactions resembling many features of cell adhesion. The process is highly specific, because vesicle docking and bilayer formation are strictly dependent on the presence of metal-affinity ligand-receptor pairs. The surface accessibility and geometry of these tethered membranes were probed by binding of histidine-tagged polypeptides. The supported membranes show adsorption kinetics and values similar to planar supported monolayers. Using various combinations of metal-chelating and histidine-tagged lipids or thiols these metal-affinity-tethered membranes should make a great impact on probing and eventually understanding the dynamic dialog of reconstituted membrane proteins.
Rädler
U
U
Cellular Biochemistry and Biophysics, Institute for Physiological Chemistry, Medical School, Philipps-University Marburg, D-35033 Marburg, Germany.
Mack
J
J
Persike
N
N
Jung
G
G
Tampé
R
R
D000345:Affinity Labels; D000595:Amino Acid Sequence; D002614:Chelating Agents; D015195:Drug Design; D006639:Histidine; D008024:Ligands; D008051:Lipid Bilayers / Q000737:chemistry*; D008670:Metals; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000737:chemistry; D013438:Sulfhydryl Compounds
D000345:Affinity Labels; D002614:Chelating Agents; D008024:Ligands; D008051:Lipid Bilayers; D008670:Metals; D010455:Peptides; D011956:Receptors, Cell Surface; D013438:Sulfhydryl Compounds; D006639:Histidine