Lipid-polymer hybrid-vesicles interrupt nucleation of amyloid fibrillation.

Solubility and aggregation of proteins are crucial factors for their functional and further biological roles. Aggregation of proteins in vivo, such as the amyloid beta (Aβ_1-40) peptide into fibrils, is significantly modulated by membrane lipids, abundantly present in cells. We developed a model membrane system, composed of lipid hybrid-vesicles bearing embedded hydrophilic polymers to in vitro study the aggregation of the Aβ_1-40 peptide. Focus is to understand and inhibit the primordial, nucleation stages of their fibrillation by added hybrid-vesicles, composed of a natural lipid and amphiphilic polymers. These designed hybrid-vesicles are based on 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC), displaying embedded hydrophilic (EO) _m P _n A_EG polymers (m = 2 or 3; P _n = 10 to 52 with M _n = 2800-9950 gmol^-1) in amounts ranging from 5-20 mol%, anchored to the POPC vesicles via hydrophobic hexadecyl-, glyceryl- and cholesteryl-moieties, affixed to the polymers as end-groups. All investigated hybrid-vesicles significantly delay fibrillation of the Aβ_1-40 peptide as determined by thioflavin T (ThT) assays. We observed that the hybrid-vesicles interacted with early aggregating species of Aβ_1-40 peptide, irrespective of their composition or size. A substantial perturbation of both primary (k ₊ k _n ) and secondary (k ₊ k ₂) nucleation rates of Aβ_1-40 by the POPC-polymer vesicles compared to POPC vesicles was observed, particularly for the cholesteryl-anchored polymers, interfering with the fragmentation and elongation steps of Aβ_1-40. Furthermore, morphological differences of the aggregates were revealed by transmission electron microscopy (TEM) images supported the inhibitory kinetic signatures.