Isolation and chemical characterization of Alzheimer's disease paired helical filament cytoskeletons

Differentiation from amyloid plaque core protein

A. E. Roher, K. C. Palmer, Vincent Chau, M. J. Ball

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

The paired helical filaments (PHFs) on Alzheimer's disease were purified by a strategy in which the neurons and amyloid plaque cores of protein (APCP) were initially isolated. This was achieved by several steps of isocratic sucrose centrifugations of increasing molarity and a discontinuous isotonic Percoll density gradient. After collagenase elimination of contaminating blood vessels, lysis of neurons was produced by SDS treatment. The released PHF cytoskeletons were separated from contaminating APCP and lipofuscin by sucrose density gradient. A final step consisted in the chemical purification of highly enriched PHFs and APCP components via a formic acid to guanidine hydrochloride transition. PHFs and APCPs were fractionated by size exclusion HPLC and further characterized and quantitated by automatic amino acid analysis. We also present some of the morphological and immunochemical characteristics of PHF polypeptides and APCP. Our studies indicate that apart from differences in localization and morphology, PHF and APCP significantly differ in (a) chemical structure (peptide and amino acid composition); (b) epitope specificity (antiubiquitin, antitau, antineurofilament); (c) physicochemical properties (structural conformation in guanidine hydrochloride); and (d) thioflavine T fluorescence emission. These parameters strongly suggest important differences in the composition and, probably, in the etiopathology of PHF and APCP of Alzheimer's disease.

Original languageEnglish (US)
Pages (from-to)2703-2716
Number of pages14
JournalJournal of Cell Biology
Volume107
Issue number6 II
StatePublished - Dec 1 1988

Fingerprint

Amyloid Plaques
Cytoskeleton
Alzheimer Disease
Proteins
formic acid
Guanidine
Sucrose
Neurons
Lipofuscin
Amino Acids
Peptides
Collagenases
Centrifugation
Blood Vessels
Epitopes
Fluorescence
High Pressure Liquid Chromatography

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

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abstract = "The paired helical filaments (PHFs) on Alzheimer's disease were purified by a strategy in which the neurons and amyloid plaque cores of protein (APCP) were initially isolated. This was achieved by several steps of isocratic sucrose centrifugations of increasing molarity and a discontinuous isotonic Percoll density gradient. After collagenase elimination of contaminating blood vessels, lysis of neurons was produced by SDS treatment. The released PHF cytoskeletons were separated from contaminating APCP and lipofuscin by sucrose density gradient. A final step consisted in the chemical purification of highly enriched PHFs and APCP components via a formic acid to guanidine hydrochloride transition. PHFs and APCPs were fractionated by size exclusion HPLC and further characterized and quantitated by automatic amino acid analysis. We also present some of the morphological and immunochemical characteristics of PHF polypeptides and APCP. Our studies indicate that apart from differences in localization and morphology, PHF and APCP significantly differ in (a) chemical structure (peptide and amino acid composition); (b) epitope specificity (antiubiquitin, antitau, antineurofilament); (c) physicochemical properties (structural conformation in guanidine hydrochloride); and (d) thioflavine T fluorescence emission. These parameters strongly suggest important differences in the composition and, probably, in the etiopathology of PHF and APCP of Alzheimer's disease.",
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Isolation and chemical characterization of Alzheimer's disease paired helical filament cytoskeletons : Differentiation from amyloid plaque core protein. / Roher, A. E.; Palmer, K. C.; Chau, Vincent; Ball, M. J.

In: Journal of Cell Biology, Vol. 107, No. 6 II, 01.12.1988, p. 2703-2716.

Research output: Contribution to journalArticle

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