Pleckstrin homology-phospholipase C-δ1 interaction with phosphatidylinositol 4,5-bisphosphate containing supported lipid bilayers monitored in situ with dual polarization interferometry.

We have determined the kinetics and affinity of binding of PH-PLCδ(1) to the PIP(2) headgroup lipids using an optical surface-sensitive technique in a time-resolved manner. The use of dual polarization interferometry to probe supported lipid bilayers (SLBs) of different compositions allowed determination of accurate affinity constants and a layer structure of the peptide binding to the model membrane platform. In addition, the platform enabled us to monitor the detailed adsorption kinetics characterized by a strong initial electrostatic attraction of the peptide to the SLB surface followed by rearrangement and loss of possibly clustered peptides upon specific binding to the phosphoinositide headgroup. These kinetics differed substantially from adsorption kinetics for nonspecific binding to similarly charged control SLBs.

Characterization of supported lipid bilayers incorporating the phosphoinositides phosphatidylinositol 4,5-biphosphate and phosphoinositol-3,4,5-triphosphate by complementary techniques.

Phosphoinositides are involved in a large number of processes in cells and it is very demanding to study individual protein-lipid interactions in vivo due to their rapid turnover and involvement in simultaneous events. Supported lipid bilayers (SLBs) containing controlled amounts of phosphoinositides provide a defined model system where important specific recognition events involving phosphoinositides can be systematically investigated using surface sensitive analytical techniques. The authors have demonstrated the formation and characterized the assembly kinetics of SLBs incorporating phosphatidylinositol 4,5-biphosphate (PIP(2); 1, 5, and 10 wt %) and phosphoinositol-3,4,5-triphosphate (1 wt %) using the quartz crystal microbalance with dissipation monitoring and fluorescence recovery after photobleaching. An increased fraction of phosphoinositides led to a higher barrier to liposome fusion, but full fluidity for the phosphatidylcholine lipids in the formed SLB. Significantly, the majority of phosphoinositides were shown to be immobile. X-ray photoelectron spectroscopy was used for the first time to verify that the PIP(2) fraction of lipids in the SLB scales linearly with the amount mixed in from stock solutions.

Understanding self-assembled amphiphilic peptide supramolecular structures from primary structure helix propensity.

Small amphiphilic peptides are attractive building blocks to design biocompatible supramolecular structures via self-assembly, with applications in, for example, drug delivery, tissue engineering, and nanotemplating. We address the influence of systematical changes in the amino acid sequence of such peptides on the self-assembled macromolecular structures. For cationic-head surfactant-like eight-residue peptides, the apolar tail amino acids were chosen to systematically vary the propensity to form an alpha-helical secondary structure while conserving the overall hydrophobicity of the sequence. Characterization of the supramolecular structures indicates that for short peptides a beta-sheet secondary structure correlates with ribbonlike assemblies while random-coil and alpha-helical secondary structures correlate with assembly of rods.