Isolation of low-molecular-weight proteins from amyloid plaque fibers in Alzheimer's disease.
During aging of the human brain, and particularly in Alzheimer's disease, progressive neuronal loss is accompanied by the formation of highly stable intra- and extraneuronal protein fibers. Using fluorescence-activated particle sorting, a method has been developed for purifying essentially to homogeneity the extracellular amyloid fibers that form the cores of senile plaques. The purified plaque cores each contain 60-130 pg of protein. Their amino acid composition shows abundant glycine, trace proline, and approximately 50% hydrophobic residues; it resembles that of enriched fractions of the paired helical filaments (PHF) that accumulate intraneuronally in Alzheimer's disease. Senile plaque amyloid fibers share with PHF insolubility in numerous protein denaturants and resistance to proteinases. However, treatment of either fiber preparation with concentrated (88%) formic acid or saturated (6.8 M) guanidine thiocyanate followed by sodium dodecyl sulfate causes disappearance of the fibers and releases proteins migrating at 5-7,000 and 11-15,000 Mr which appear to be dimerically related. Following their separation by size-exclusion HPLC, the proteins solubilized from plaque amyloid and PHF-enriched fractions have highly similar compositions and, on dialysis, readily aggregate into higher Mr polymers. Antibodies raised to the major low-Mr protein selectively label both plaque cores and vascular amyloid deposits in Alzheimer brain but do not stain neurofibrillary tangles, senile plaque neurites, or any other neuronal structure. Thus, extraneuronal amyloid plaque filaments in Alzheimer's disease are composed of hydrophobic low-Mr protein(s) which are also present in vascular amyloid deposits. Current evidence suggests that such protein(s) found in PHF-enriched fractions may derive from copurifying amyloid filaments rather than from PHF.
D000368:Aged; D000544:Alzheimer Disease / Q000473:pathology*; D000596:Amino Acids / Q000032:analysis; D000682:Amyloid / Q000032:analysis*; D001921:Brain / Q000473:pathology; D002851:Chromatography, High Pressure Liquid; D005434:Flow Cytometry; D006651:Histocytochemistry; D006801:Humans; D007163:Immunosorbent Techniques; D008854:Microscopy, Electron; D008875:Middle Aged; D008970:Molecular Weight; D009454:Neurofibrils / Q000032:analysis; D011506:Proteins / Q000032:analysis*
D000596:Amino Acids; D000682:Amyloid; D011506:Proteins