Analysis of single Alzheimer solid plaque cores by laser capture microscopy and nanoelectrospray/tandem mass spectrometry.
Aggregation of the 40-42 residue amyloid beta-peptide (Abeta) into amyloid plaques is a central event in Alzheimer's disease (AD) pathogenesis. Many proteins have by immunohistochemical techniques been shown to codeposit with Abeta in AD plaques. It is possible that some of these could seed Abeta aggregation and therefore be found in the actual core of the plaque. Here, we present a highly sensitive method for unbiased biochemical analysis of plaque cores. A mild purification protocol based on centrifugation and filtration was used to purify intact plaque cores from human AD brain. The purified plaques were dispensed on a glass slide and viewed in a laser capture microscope, and plaque cores were catapulted into a tube cap by a laser beam. After dissolution in formic acid, plaques were digested and analyzed by liquid chromatography coupled online to electrospray/tandem mass spectrometry. One single plaque was found to be sufficient for positive identification of the main amyloid component. Remarkably, Abeta was the only protein identified when 200 plaques were isolated and analyzed with the present method. Thus, it is possible that no proteins copolymerize with Abeta in the plaque cores and that Abeta alone is sufficient for formation of plaque cores. In support of this notion, core-like structures were observed after incubation of synthetic Abeta for 2 weeks. We suggest that the method described here could be used for the general analysis of amyloid aggregates and inclusion bodies found in other neurodegenerative disorders and that plaque cores in AD brain are molecularly homogeneous structures.
Söderberg
Linda
L
Karolinska Institutet and Dainippon Sumitomo Pharma Alzheimer Center (KASPAC), Neurotec, Novum.
Bogdanovic
Nenad
N
Axelsson
Birgitta
B
Winblad
Bengt
B
Näslund
Jan
J
Tjernberg
Lars O
LO
D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000473:pathology*; D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D017809:Fatal Outcome; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D058225:Plaque, Amyloid / Q000737:chemistry / Q000473:pathology*; D020836:Protein Structure, Quaternary; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D016229:Amyloid beta-Peptides