Alzheimer's beta-peptide oligomer formation at physiologic concentrations.
When diluted from dimethyl sulfoxide or 1,1,1,3,3,3-hexafluoro-2-propanol, synthetic human Abeta(1-42) readily forms oligomeric structures at near physiologic concentrations (1-20 nM). Oligomers 40 kDa are detected in a sandwich enzyme-linked immunosorbant assay where the capture and detection antibodies recognize the same primary sequence epitope. Monomeric peptide with a single epitope does not react in this format. Abeta(1-40) peptide does not oligomerize readily under these conditions. The rate of oligomer formation has a steep linear temperature dependence but is weakly affected by ionic strength up to 0.5M NaCl or KCl. Oligomer formation is inhibited by concentrations of Tween 20 and several other detergents well below their critical micelle concentrations. Once formed, high-molecular-weight oligomers are stabilized by Tween 20. Gel permeation chromatography of an oligomer preparation formed at nanomolar concentrations indicates that the majority of the Abeta(1-42) peptide chromatographs as monomers/dimers of apparent mw approximately 10 kDa. The most abundant oligomers have apparent mobilities corresponding to 220 kDa (48-mer) and higher multiples of this without detectable concentrations of intermediate low-molecular-weight species. Very little immunoreactive peptide appears in the void volume (>1.5 MDa) of a Superose 12 column. The oligomers are stable, rechromatographing at their original position. Abeta(1-42) oligomer formation at physiologic concentrations is a reproducible process that is amenable to kinetic analysis and inhibition.
LeVine
Harry
H
Department of Molecular and Cellular Biochemistry, Chandler School of Medicine and the Center on Aging, University of Kentucky, Lexington, KY 40536-0230, USA. hlevine@email.uky.edu
D000544:Alzheimer Disease / Q000378:metabolism* / Q000473:pathology; D016229:Amyloid beta-Peptides / Q000302:isolation & purification / Q000378:metabolism*; D015153:Blotting, Western; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents / Q000494:pharmacology; D019281:Dimerization; D006801:Humans; D008027:Light; D008823:Micelles; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism*; D012680:Sensitivity and Specificity
D016229:Amyloid beta-Peptides; D003432:Cross-Linking Reagents; D008823:Micelles; D010446:Peptide Fragments; C078592:amyloid beta-protein (1-40); C075222:amyloid beta-protein (1-42)