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Preparation of Pure Populations of Amyloid β-Protein Oligomers of Defined Size. | LitMetric

Preparation of Pure Populations of Amyloid β-Protein Oligomers of Defined Size.

Methods Mol Biol

Department of Neurology, Molecular Biology Institute, Brain Research Institute, David Geffen School of Medicine at UCLA, University of California, Los Angeles, Los Angeles, CA, USA.

Published: July 2019

AI Article Synopsis

Article Abstract

Protein and peptide oligomers are thought to play important roles in the pathogenesis of a number of neurodegenerative diseases. For this reason, considerable effort has been devoted to understanding the oligomerization process and to determining structure-activity relationships among the many types of oligomers that have been described. We discuss here a method for producing pure populations of amyloid β-protein (Aβ) of specific sizes using the most pathologic form of the peptide, Aβ42. This work was necessitated because Aβ oligomerization produces oligomers of many different sizes that are non-covalently associated, which means that dissociation or further assembly may occur. These characteristics preclude rigorous structure-activity determinations. In studies of Aβ40, we have used the method of photo-induced cross-linking of unmodified proteins (PICUP) to produce zero-length carbon-carbon bonds among the monomers comprising each oligomer, thus stabilizing the oligomers. We then isolated pure populations of oligomers by fractionating the oligomers by size using SDS-PAGE and then extracting each population from the stained gel bands. Although this procedure worked well with the shorter Aβ40 peptide, we found that a significant percentage of Aβ42 oligomers had not been stabilized. Here, we discuss a new method capable of yielding stable Aβ42 oligomers of sizes from dimer through dodecamer.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6490168PMC
http://dx.doi.org/10.1007/978-1-4939-7816-8_1DOI Listing

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