Influence of the Human and Rat Islet Amyloid Polypeptides on Structure of Phospholipid Bilayers: Neutron Reflectometry and Fluorescence Microscopy Studies.

Neutron reflectivity (NR) and fluorescent microscopy (FM) were used to study the interactions of human (hIAPP) and rat (rIAPP) islet amyloid polypeptides with several formulations of supported model lipid bilayers at the solid-liquid interface. Aggregation and deposition of islet amyloid polypeptide is correlated with the pathology of many diseases, including Alzheimer's, Parkinson, and type II diabetes (T2DM). A central component of T2DM pathology is the deposition of fibrils in the endocrine pancreas, which is toxic to the insulin secreting ß-cells. The molecular mechanism by which the cell death occurs is not yet understood, but existing evidence points toward interactions of IAPP oligomers with cellular membranes in a manner leading to loss of their integrity. Our NR and FM results showed that the human sequence variant, hIAPP, had little or no effect on bilayers composed of saturated-acyl chains like zwitterionic DPPC, anionic DPPG, and mixed 80:20 mol % DPPC:DPPG bilayers. In marked contrast, the bilayer structure and stability of anionic unsaturated DOPG were sensitive to protein interaction, and the bilayer was partly solubilized by hIAPP under the conditions used here. The rIAPP, which is considered less toxic, had no perturbing effects on any of the above membrane formulations. Understanding the conditions that result in membrane disruption by hIAPP can be crucial in developing counter strategies to fight T2DM and also physicochemically similar neurodegenerative diseases such as Alzheimer's.

A unified model of protein dynamics.

Protein functions require conformational motions. We show here that the dominant conformational motions are slaved by the hydration shell and the bulk solvent. The protein contributes the structure necessary for function. We formulate a model that is based on experiments, insights from the physics of glass-forming liquids, and the concepts of a hierarchically organized energy landscape. To explore the effect of external fluctuations on protein dynamics, we measure the fluctuations in the bulk solvent and the hydration shell with broadband dielectric spectroscopy and compare them with internal fluctuations measured with the Mössbauer effect and neutron scattering. The result is clear. Large-scale protein motions are slaved to the fluctuations in the bulk solvent. They are controlled by the solvent viscosity, and are absent in a solid environment. Internal protein motions are slaved to the beta fluctuations of the hydration shell, are controlled by hydration, and are absent in a dehydrated protein. The model quantitatively predicts the rapid increase of the mean-square displacement above approximately 200 K, shows that the external beta fluctuations determine the temperature- and time-dependence of the passage of carbon monoxide through myoglobin, and explains the nonexponential time dependence of the protein relaxation after photodissociation.