sodium-dodecyl-sulfate and Dementia

Soluble non-fibrillar assemblies of amyloid-beta (Aβ) and aggregated tau protein are the proximate synaptotoxic species associated with Alzheimer's disease (AD). Anti-Aβ immunotherapy is a promising and advanced therapeutic strategy, but the precise Aβ species to target is not yet known. Previously, we and others have shown that natural human IgGs (NAbs) target diverse Aβ conformers and have therapeutic potential. We now demonstrate that these antibodies bound with nM avidity to conformational epitopes on plate-immobilized synthetic Aβ dimer assemblies, including synaptotoxic protofibrils, and targeted these conformers in solution. Importantly, NAbs also recognized Aβ extracted from the water-soluble phase of human AD brain, including species that migrated on denaturing PAGE as SDS-stable dimers. The critical reliance on Aβ's conformational state for NAb binding, and not a linear sequence epitope, was confirmed by the antibody's nM reactivity with plate-immobilized protofibrills, and weak uM binding to synthetic Aβ monomers and peptide fragments. The antibody's lack of reactivity against a linear sequence epitope was confirmed by our ability to isolate anti-Aβ NAbs from intravenous immunoglobulin using affinity matrices, immunoglobulin light chain fibrils and Cibacron blue, which had no sequence similarity with the peptide. These findings suggest that further investigations on the molecular basis and the therapeutic/diagnostic potential of anti-Aβ NAbs are warranted.

Topics: Aged; Amyloid beta-Peptides; Benzothiazoles; Biophysics; Brain; Circular Dichroism; Dementia; Dimerization; Electrophoresis, Polyacrylamide Gel; Epitopes; Female; Humans; Immunoglobulins; Immunoglobulins, Intravenous; Microscopy, Electron; Middle Aged; Peptides; Protein Conformation; Sodium Dodecyl Sulfate; Thiazoles

The molecular pathways leading to Alzheimer-type dementia are not well understood, but the amyloid beta-protein is believed to be centrally involved. The quantity of amyloid beta-protein containing plaques does not correlate well with clinical status, suggesting that if amyloid beta-protein is pathogenic it involves soluble non-plaque material. Using 43 brains from the Newcastle cohort of the population-representative Medical Research Council Cognitive Function and Ageing Study, we examined the relationship between biochemically distinct forms of amyloid beta-protein and the presence of Alzheimer-type dementia. Cortical samples were serially extracted with Tris-buffered saline, Tris-buffered saline containing 1% TX-100 and with 88% formic acid and extracts analysed for amyloid beta-protein by immunoprecipitation/western blotting. The cohort was divisible into those with dementia at death with (n = 14) or without (n = 10) significant Alzheimer-type pathology, and those who were not demented (n = 19). Amyloid beta-protein monomer in extracts produced using Tris-buffered saline and Tris-buffered saline containing 1% TX-100 were strongly associated with Alzheimer type dementia (P < 0.001) and sodium dodecyl sulphate-stable amyloid beta-protein dimer was detected specifically and sensitively in Tris-buffered saline, Tris-buffered saline containing 1% TX-100 and formic acid extracts of Alzheimer brain. Amyloid beta-protein monomer in the formic acid fraction closely correlated with diffuse and neuritic plaque burden, but was not specific for dementia. These findings support the hypothesis that soluble amyloid beta-protein is a major correlate of dementia associated with Alzheimer-type pathology and is likely to be intimately involved in the pathogenesis of cognitive failure.

Topics: Aged; Aged, 80 and over; Alzheimer Disease; Amyloid beta-Peptides; Blotting, Western; Cerebral Cortex; Cohort Studies; Dementia; Drug Stability; Female; Humans; Immunoprecipitation; Male; Protein Multimerization; Sodium Dodecyl Sulfate

In recent years, several attempts have been made to identify biochemical and/or genetic markers which might have diagnostic and prognostic uses for Alzheimer's disease (AD). To look for possible new blood markers, a longitudinal study was carried out in our Central Nervous System Biomedical Research Center between October 1996 and July 1998. A total of 30 AD subjects were diagnosed as AD patients according to the DSM-IV and NINCDS-ADRDA criteria. Vascular dementia (VD, n= 17), mixed dementia (MXD, n = 18) and healthy age-matched control subjects (n = 15) have been included in the study. Serum samples have been analysed by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and the resulting protein patterns have been compared. The objectives of this study were to examine the presence of peripheral markers among patients with AD, VD and MXD and to explore the relationship between serum markers and APOE genotype. The findings suggest that two proteins of 19 and 20kDa molecular weight, respectively, might be associated with disease progression in different types of dementia.

Topics: Aged; Aged, 80 and over; Alzheimer Disease; Biomarkers; Blood Proteins; Dementia; Dementia, Vascular; Electrophoresis, Polyacrylamide Gel; Humans; Middle Aged; Prognosis; Sodium Dodecyl Sulfate

Topics: Acyltransferases; Alzheimer Disease; Dementia; Guanidine; Guanidines; Humans; Neurofibrils; Octoxynol; Polyethylene Glycols; Sodium Dodecyl Sulfate; Solubility; Transglutaminases; Urea

A method is described for the partial purification of the paired helical filaments that accumulate progressively in human neurons in Alzheimer's disease (senile dementia). Paired helical filaments have unusual solubility characteristics, including insolubility in sodium dodecyl sulfate, urea, reducing agent, and guanidine, which prevent analysis of their molecular composition by gel electrophoresis. The paired helical filaments appear to contain covalent bonds other than disulfide, which cross-link individual filaments into a rigid intracellular polymer. Thus, paired helical filaments appear to represent an example in neurons of an insoluble cross-linked protein. Covalently cross-linked protein polymers occur in lens senile cataracts and in terminally differentiated skin keratinocytes, suggesting that there may be a common mechanism for remodeling some structural proteins during cell aging.

Topics: Alzheimer Disease; Cerebral Cortex; Cytoskeleton; Dementia; gamma-Glutamyltransferase; Humans; Microscopy, Electron; Sodium Dodecyl Sulfate; Solubility; Urea

Topics: Dementia; Electrophoresis, Polyacrylamide Gel; Hippocampus; Humans; Methods; Microscopy, Electron; Microscopy, Phase-Contrast; Molecular Weight; Nerve Tissue Proteins; Parkinson Disease; Sodium Dodecyl Sulfate; Spectrum Analysis