Title
Amyloid-β 1-42 Aggregation Initiates Its Cellular Uptake and Cytotoxicity
Author
Sha Jin, Niraja Kedia, Eva Illes-Toth, Ivan Haralampiev, Simon Prisner, Andreas Herrmann, Erich E. Wanker, Jan Bieschke
Year
2016
Journal
Journal of Biological Chemistry
Abstract
The accumulation of Amyloid beta peptide1-42 (Aβ1-42) in extracellular plaques is one of the pathological hallmarks of Alzheimer's disease (AD). Several studies have suggested that cellular reuptake of Aβ1-42 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 Aβ1-42 and tracked its internalization by human neuroblastoma cells and neurons. β-sheet-rich Aβ1-42 aggregates entered the cells at low nanomolar concentration of Aβ1-42. In contrast, monomer uptake faced a concentration threshold and occurred only at concentrations and time scales that allowed Aβ1-42 aggregates to form. By uncoupling membrane binding from internalization, we found that Aβ1-42 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 Aβ1-42 uptake and cytotoxicity.
Instrument
J-720
Keywords
Circular dichroism, Secondary structure, Aggregation, Biochemistry