Amyloid beta-peptide polymerization studied using fluorescence correlation spectroscopy.
BACKGROUND
The accumulation of fibrillar deposits of amyloid beta-peptide (Abeta) in brain parenchyma and cerebromeningeal blood vessels is a key step in the pathogenesis of Alzheimer's disease. In this report, polymerization of Abeta was studied using fluorescence correlation spectroscopy (FCS), a technique capable of detecting small molecules and large aggregates simultaneously in solution.


RESULTS
The polymerization of Abeta dissolved in Tris-buffered saline, pH 7.4, occurred above a critical concentration of 50 microM and proceeded from monomers/dimers into two discrete populations of large aggregates, without any detectable amount of oligomers. The aggregation showed very high cooperativity and reached a maximum after 40 min, followed by an increase in the amount of monomers/dimers and a decrease in the size of the large aggregates. Electron micrographs of samples prepared at the time for maximum aggregation showed a mixture of an amorphous network and short diffuse fibrils, whereas only mature amyloid fibrils were detected after one day of incubation. The aggregation was reduced when Abeta was incubated in the presence of Abeta ligands, oligopeptides previously shown to inhibit fibril formation, and aggregates were partly dissociated after the addition of the ligands.


CONCLUSIONS
The polymerization of Abeta is a highly cooperative process in which the formation of very large aggregates precedes the formation of fibrils. The entire process can be inhibited and, at least in early stages, partly reversed by Abeta ligands.
Tjernberg
L O
LO
Laboratory of Biochemistry and Molecular Pharmacology, Section of Drug Dependence Research, Department of Clinical Neuroscience, the Karolinska Hospital, Stockholm, Sweden. Lars.Tjernberg@cmm.ki.se
Pramanik
A
A
Björling
S
S
Thyberg
P
P
Thyberg
J
J
Nordstedt
C
C
Berndt
K D
KD
Terenius
L
L
Rigler
R
R
D000544:Alzheimer Disease / Q000378:metabolism; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D001704:Biopolymers / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D005456:Fluorescent Dyes; D006801:Humans; D008024:Ligands; D008854:Microscopy, Electron; D010455:Peptides / Q000032:analysis / Q000378:metabolism; D012235:Rhodamines; D013050:Spectrometry, Fluorescence / Q000379:methods
D016229:Amyloid beta-Peptides; D001704:Biopolymers; D005456:Fluorescent Dyes; D008024:Ligands; D010455:Peptides; D012235:Rhodamines