title: Polyglutamine aggregates impair lipid membrane integrity and enhance lipid membrane rigidity.
abstract: Lipid membranes are suggested as the primary target of amyloid aggregates. We study aggregates formed by a polyglutamine (polyQ) peptide, and their disruptive effect on lipid membranes. Using solution atomic force microscopy (AFM), we observe polyQ oligomers coexisting with short fibrils, which have a twisted morphology that likely corresponds to two intertwined oligomer strings. Fourier transform infrared spectroscopy reveals that the content of β-sheet enriched aggregates increases with incubation time. Using fluorescence microscopy, we find that exposure to polyQ aggregates results in deflated morphology of giant unilamellar vesicles. PolyQ aggregates induced membrane disruption is further substantiated by time-dependent calcein leakage from the interior to the exterior of lipid vesicles. Detailed structural and mechanical perturbations of lipid membranes are revealed by solution AFM. We find that membrane disruption by polyQ aggregates proceeds by a two-step process, involving partial and full disruption. In addition to height contrast, the resulting partially and fully disrupted bilayers have distinct rigidity and adhesion force properties compared to the intact bilayer. Specifically, the bilayer rigidity increases as the intact bilayer becomes partially and fully disrupted. Surprisingly, the adhesion force first decreases and then increases during the disruption process. By resolving individual fibrils deposited on bilayer surface, we show that both the length and the number of fibrils can increase with incubation time. Our results highlight that membrane disruption could be the molecular basis of polyQ aggregates induced cytotoxicity.
authors.affiliation: Department of Physics, University of South Florida, Tampa, FL 33620, United States.
authors.forename: Chian Sing
authors.initials: CS
authors.lastname: Ho
authors.affiliation: Department of Physics, University of South Florida, Tampa, FL 33620, United States.
authors.forename: Nawal K
authors.initials: NK
authors.lastname: Khadka
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, FL 33620, United States.
authors.forename: Fengyu
authors.initials: F
authors.lastname: She
authors.affiliation: Department of Chemistry, University of South Florida, Tampa, FL 33620, United States.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Cai
authors.affiliation: Department of Physics, University of South Florida, Tampa, FL 33620, United States. Electronic address: panj@usf.edu.
authors.forename: Jianjun
authors.initials: J
authors.lastname: Pan
mesh_terms: D000682:Amyloid / Q000737:chemistry / Q000378:metabolism*; D058227:Amyloidogenic Proteins / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D018625:Microscopy, Atomic Force; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D066263:Protein Aggregation, Pathological / Q000378:metabolism*; D053835:Unilamellar Liposomes / Q000737:chemistry
chemical_list: D000682:Amyloid; D058227:Amyloidogenic Proteins; D008051:Lipid Bilayers; D010455:Peptides; D053835:Unilamellar Liposomes; C097188:polyglutamine
keywords: Amyloid; Fluorescence microscopy; Lipid bilayer; Oligomer; Polyglutamine; Solution AFM