Morphological and biochemical analyses of amyloid plaque core proteins purified from Alzheimer disease brain tissue.
Amyloid plaque cores were purified from Alzheimer disease brain tissue. Plaque core proteins were solubilized in formic acid which upon dialysis against guanidinium hydrochloride (GuHCl) partitioned into soluble (approximately 15%) and insoluble (approximately 85%) components. The GuHCl-soluble fraction contained beta-amyloid1-40, whereas the GuHCl-insoluble fraction was fractionated into six components by size exclusion HPLC: S1 (> 200 kDa), S2 (200 kDa), S3 (45 kDa), S4 (15 kDa), S5 (10 kDa), and S6 (5 kDa). Removal of the GuHCl reconstituted 10-nm filaments composed of two intertwined 5-nm strands. Fractions S5 and S6 also yielded filamentous structures when treated similarly, whereas fractions S1-S4 yielded amorphous aggregates. Chemical analysis identified S4-S6 as multimeric and monomeric beta-amyloid. Immunochemical analyses revealed alpha 1-antichymotrypsin and non-beta-amyloid segments of the beta-amyloid precursor protein within fractions S1 and S2. Several saccharide components were identified within plaque core protein preparations by fluorescence and electron microscopy, as seen with fluorescein isothiocyanate- and colloidal gold-conjugated lectins. We have shown previously that this plaque core protein complex is more toxic to neuronal cultures than beta-amyloid. The non-beta-amyloid components likely mediate this additional toxicity, imposing a significant influence on the pathophysiology of Alzheimer disease.
Roher
A E
AE
Department of Anatomy and Cell Biology, Wayne State University School of Medicine, Detroit, MI 48201.
Palmer
K C
KC
Yurewicz
E C
EC
Ball
M J
MJ
Greenberg
B D
BD
D000368:Aged; D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000378:metabolism / Q000473:pathology*; D000596:Amino Acids / Q000032:analysis; D000682:Amyloid / Q000032:analysis* / Q000737:chemistry* / Q000648:ultrastructure; D016229:Amyloid beta-Peptides / Q000032:analysis / Q000737:chemistry* / Q000648:ultrastructure; D016564:Amyloid beta-Protein Precursor / Q000032:analysis / Q000737:chemistry; D001921:Brain / Q000473:pathology* / Q000648:ultrastructure; D002241:Carbohydrates / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D037102:Lectins; D046911:Macromolecular Substances; D008297:Male; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron
D000596:Amino Acids; D000682:Amyloid; D016229:Amyloid beta-Peptides; D016564:Amyloid beta-Protein Precursor; D002241:Carbohydrates; D037102:Lectins; D046911:Macromolecular Substances