title: Amyloid-β(1-42) Aggregation Initiates Its Cellular Uptake and Cytotoxicity.
abstract: The accumulation of amyloid β peptide(1-42) (Aβ(1-42)) in extracellular plaques is one of the pathological hallmarks of Alzheimer disease (AD). Several studies have suggested that cellular reuptake of Aβ(1-42) may be a crucial step in its cytotoxicity, but the uptake mechanism is not yet understood. Aβ may be present in an aggregated form prior to cellular uptake. Alternatively, monomeric peptide may enter the endocytic pathway and conditions in the endocytic compartments may induce the aggregation process. Our study aims to answer the question whether aggregate formation is a prerequisite or a consequence of Aβ endocytosis. We visualized aggregate formation of fluorescently labeled Aβ(1-42) and tracked its internalization by human neuroblastoma cells and neurons. β-Sheet-rich Aβ(1-42) aggregates entered the cells at low nanomolar concentration of Aβ(1-42). In contrast, monomer uptake faced a concentration threshold and occurred only at concentrations and time scales that allowed Aβ(1-42) aggregates to form. By uncoupling membrane binding from internalization, we found that Aβ(1-42) monomers bound rapidly to the plasma membrane and formed aggregates there. These structures were subsequently taken up and accumulated in endocytic vesicles. This process correlated with metabolic inhibition. Our data therefore imply that the formation of β-sheet-rich aggregates is a prerequisite for Aβ(1-42) uptake and cytotoxicity.
authors.affiliation: From the Proteomics and Molecular Mechanisms of Neurodegenerative Diseases, Max Delbrück Center for Molecular Medicine, 13125 Berlin-Buch, Germany, the Department of Biomedical Engineering, Washington University, St. Louis, Missouri 63130.
authors.forename: Sha
authors.initials: S
authors.lastname: Jin
authors.affiliation: the Department of Biomedical Engineering, Washington University, St. Louis, Missouri 63130.
authors.forename: Niraja
authors.initials: N
authors.lastname: Kedia
authors.affiliation: the Department of Biomedical Engineering, Washington University, St. Louis, Missouri 63130.
authors.forename: Eva
authors.initials: E
authors.lastname: Illes-Toth
authors.affiliation: the Department of Biology and.
authors.forename: Ivan
authors.initials: I
authors.lastname: Haralampiev
authors.affiliation: the Department of Biology and.
authors.forename: Simon
authors.initials: S
authors.lastname: Prisner
authors.affiliation: IRI Life Sciences, Humboldt-Universität zu Berlin, 10115 Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Herrmann
authors.affiliation: From the Proteomics and Molecular Mechanisms of Neurodegenerative Diseases, Max Delbrück Center for Molecular Medicine, 13125 Berlin-Buch, Germany.
authors.forename: Erich E
authors.initials: EE
authors.lastname: Wanker
authors.affiliation: the Department of Biomedical Engineering, Washington University, St. Louis, Missouri 63130, bieschke@wustl.edu.
authors.forename: Jan
authors.initials: J
authors.lastname: Bieschke
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism* / Q000473:pathology; D016229:Amyloid beta-Peptides / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism* / Q000473:pathology; D004705:Endocytosis; D006801:Humans; D010446:Peptide Fragments / Q000378:metabolism*; D066263:Protein Aggregation, Pathological / Q000378:metabolism* / Q000473:pathology; D017433:Protein Structure, Secondary; D021381:Protein Transport
chemical_list: D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; C075222:amyloid beta-protein (1-42)
keywords: aggregation; amyloid; amyloid-β (AB); internalization; kinetics; protein folding