Preparation and characterization of toxic Abeta aggregates for structural and functional studies in Alzheimer's disease research.

Asad Jan, Dean M. Hartley, Hilal A. Lashuel

Research output: Contribution to journalArticle

153 Citations (Scopus)

Abstract

The amyloid cascade hypothesis, supported by strong evidence from genetics, pathology and studies using animal models, implicates amyloid-beta (Abeta) oligomerization and fibrillogenesis as central causative events in the pathogenesis of Alzheimer's disease (AD). Today, significant efforts in academia, biotechnology and the pharmaceutical industry are devoted to identifying the mechanisms by which the process of Abeta aggregation contributes to neurodegeneration in AD and to the identity of the toxic Abeta species. In this paper, we describe methods and detailed protocols for reproducibly preparing Abeta aggregates of defined size distribution and morphology, including monomers, protofibrils and fibrils, using size exclusion chromatography. In addition, we describe detailed biophysical procedures for elucidating the structural features, aggregation kinetics and toxic properties of the different Abeta aggregation states, with special emphasis on protofibrillar intermediates. The information provided by this approach allows for consistent correlation between the properties of the aggregates and their toxicity toward primary neurons and/or cell lines. A better understanding of the molecular and structural basis of Abeta aggregation and toxicity is crucial for the development of effective strategies aimed at prevention and/or treatment of AD. Furthermore, the identification of specific aggregation states, which correlate with neurodegeneration in AD, could lead to the development of diagnostic tools to detect and monitor disease progression. The procedures described can be performed in as little as 1 day, or may take longer, depending on the exact toxicity assays used.

Original languageEnglish
Pages (from-to)1186-1209
Number of pages24
JournalNature Protocols
Volume5
Issue number6
DOIs
Publication statusPublished - 22 Sep 2010
Externally publishedYes

Fingerprint

Poisons
Amyloid
Alzheimer Disease
Agglomeration
Research
toxicity
Toxicity
pharmaceutical industry
biotechnology
pathology
chromatography
Oligomerization
Size exclusion chromatography
Drug Industry
Pathology
Biotechnology
assay
Neurons
Gel Chromatography
kinetics

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Preparation and characterization of toxic Abeta aggregates for structural and functional studies in Alzheimer's disease research. / Jan, Asad; Hartley, Dean M.; Lashuel, Hilal A.

In: Nature Protocols, Vol. 5, No. 6, 22.09.2010, p. 1186-1209.

Research output: Contribution to journalArticle

@article{5af628c184bf475eb0bf2a388f669a29,
title = "Preparation and characterization of toxic Abeta aggregates for structural and functional studies in Alzheimer's disease research.",
abstract = "The amyloid cascade hypothesis, supported by strong evidence from genetics, pathology and studies using animal models, implicates amyloid-beta (Abeta) oligomerization and fibrillogenesis as central causative events in the pathogenesis of Alzheimer's disease (AD). Today, significant efforts in academia, biotechnology and the pharmaceutical industry are devoted to identifying the mechanisms by which the process of Abeta aggregation contributes to neurodegeneration in AD and to the identity of the toxic Abeta species. In this paper, we describe methods and detailed protocols for reproducibly preparing Abeta aggregates of defined size distribution and morphology, including monomers, protofibrils and fibrils, using size exclusion chromatography. In addition, we describe detailed biophysical procedures for elucidating the structural features, aggregation kinetics and toxic properties of the different Abeta aggregation states, with special emphasis on protofibrillar intermediates. The information provided by this approach allows for consistent correlation between the properties of the aggregates and their toxicity toward primary neurons and/or cell lines. A better understanding of the molecular and structural basis of Abeta aggregation and toxicity is crucial for the development of effective strategies aimed at prevention and/or treatment of AD. Furthermore, the identification of specific aggregation states, which correlate with neurodegeneration in AD, could lead to the development of diagnostic tools to detect and monitor disease progression. The procedures described can be performed in as little as 1 day, or may take longer, depending on the exact toxicity assays used.",
author = "Asad Jan and Hartley, {Dean M.} and Lashuel, {Hilal A.}",
year = "2010",
month = "9",
day = "22",
doi = "10.1038/nprot.2010.72",
language = "English",
volume = "5",
pages = "1186--1209",
journal = "JAPCA",
issn = "1073-161X",
publisher = "Taylor and Francis Ltd.",
number = "6",

}

TY - JOUR

T1 - Preparation and characterization of toxic Abeta aggregates for structural and functional studies in Alzheimer's disease research.

AU - Jan, Asad

AU - Hartley, Dean M.

AU - Lashuel, Hilal A.

PY - 2010/9/22

Y1 - 2010/9/22

N2 - The amyloid cascade hypothesis, supported by strong evidence from genetics, pathology and studies using animal models, implicates amyloid-beta (Abeta) oligomerization and fibrillogenesis as central causative events in the pathogenesis of Alzheimer's disease (AD). Today, significant efforts in academia, biotechnology and the pharmaceutical industry are devoted to identifying the mechanisms by which the process of Abeta aggregation contributes to neurodegeneration in AD and to the identity of the toxic Abeta species. In this paper, we describe methods and detailed protocols for reproducibly preparing Abeta aggregates of defined size distribution and morphology, including monomers, protofibrils and fibrils, using size exclusion chromatography. In addition, we describe detailed biophysical procedures for elucidating the structural features, aggregation kinetics and toxic properties of the different Abeta aggregation states, with special emphasis on protofibrillar intermediates. The information provided by this approach allows for consistent correlation between the properties of the aggregates and their toxicity toward primary neurons and/or cell lines. A better understanding of the molecular and structural basis of Abeta aggregation and toxicity is crucial for the development of effective strategies aimed at prevention and/or treatment of AD. Furthermore, the identification of specific aggregation states, which correlate with neurodegeneration in AD, could lead to the development of diagnostic tools to detect and monitor disease progression. The procedures described can be performed in as little as 1 day, or may take longer, depending on the exact toxicity assays used.

AB - The amyloid cascade hypothesis, supported by strong evidence from genetics, pathology and studies using animal models, implicates amyloid-beta (Abeta) oligomerization and fibrillogenesis as central causative events in the pathogenesis of Alzheimer's disease (AD). Today, significant efforts in academia, biotechnology and the pharmaceutical industry are devoted to identifying the mechanisms by which the process of Abeta aggregation contributes to neurodegeneration in AD and to the identity of the toxic Abeta species. In this paper, we describe methods and detailed protocols for reproducibly preparing Abeta aggregates of defined size distribution and morphology, including monomers, protofibrils and fibrils, using size exclusion chromatography. In addition, we describe detailed biophysical procedures for elucidating the structural features, aggregation kinetics and toxic properties of the different Abeta aggregation states, with special emphasis on protofibrillar intermediates. The information provided by this approach allows for consistent correlation between the properties of the aggregates and their toxicity toward primary neurons and/or cell lines. A better understanding of the molecular and structural basis of Abeta aggregation and toxicity is crucial for the development of effective strategies aimed at prevention and/or treatment of AD. Furthermore, the identification of specific aggregation states, which correlate with neurodegeneration in AD, could lead to the development of diagnostic tools to detect and monitor disease progression. The procedures described can be performed in as little as 1 day, or may take longer, depending on the exact toxicity assays used.

UR - http://www.scopus.com/inward/record.url?scp=77956698237&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77956698237&partnerID=8YFLogxK

U2 - 10.1038/nprot.2010.72

DO - 10.1038/nprot.2010.72

M3 - Article

VL - 5

SP - 1186

EP - 1209

JO - JAPCA

JF - JAPCA

SN - 1073-161X

IS - 6

ER -