Graphical Analysis of Mass and Anisotropy Changes Observed by Plasmon-Waveguide Resonance Spectroscopy Can Provide Useful Insights into Membrane Protein Function

Plasmon-waveguide resonance spectroscopy is a recently developed optical method that allows characterization of mass and structural changes in two-dimensionally ordered thin films (e.g., proteolipid membranes) deposited onto a sensor surface. Full analysis of these systems involves fitting theoretical curves (obtained using Maxwell's equations) to experimental spectra measured using s- and p-polarized excitation. This allows values to be obtained for refractive indices and optical extinction coefficients in these two directions, as well as a value for film thickness, thereby providing information about mass density and anisotropy changes. This is a time-consuming process that works well for simple systems in which only a single conformational event occurs, but cannot distinguish between events involving multiple conformations that proceed either sequentially or in a parallel series of events. This article describes a graphical method that can distinguish between mass density and anisotropy changes in a simpler, more rapid procedure, even for processes that proceed via multiple conformational events. This involves measurement of plasmon-waveguide resonance spectral shifts obtained upon molecular interactions occurring in deposited films with both s- and p-polarized excitation, and transforming these from an ( s- p) coordinate system into a (mass-structure) coordinate system. This procedure is illustrated by data obtained upon the binding of a small peptide, penetratin, to solid-supported lipid bilayer membranes.