In vitro characterization of conditions for amyloid-β peptide oligomerization and fibrillogenesis
Open accessAbstract
Extensive research causally links amyloid-beta peptide (A beta) to Alzheimer's disease, although the pathologically relevant A beta conformation remains unclear. A beta spontaneously aggregates into the fibrils that deposit in senile plaques. However, recent in vivo and in vitro reports describe a potent biological activity for oligomeric assemblies of A beta. To consistently prepare in vitro oligomeric and fibrillar forms of A beta 1-42, a detailed knowledge of how solution parameters influence structure is required. This manuscript represents the first study using a single chemically and structurally homogeneous unaggregated starting material to demonstrate that the formation of oligomers, fibrils, and fibrillar aggregates is determined by time, concentration, temperature, pH, ionic strength, and A beta species. We recently reported that oligomers inhibit neuronal viability 10-fold more than fibrils and approximately 40-fold more than unaggregated peptide, with oligomeric A beta 1-42-induced neurotoxicity significant at 10 nm. In addition, we were able to differentiate by structure and neurotoxic activity wild-type A beta1-42 from isoforms containing familial mutations (Dahlgren, K. N., Manelli, A. M., Stine, W. B., Jr., Baker, L. K., Krafft, G. A., and LaDu, M. J. (2002) J. Biol. Chem. 277, 32046-32053). Understanding the biological role of specific A beta conformations may define the link between A beta and Alzheimer's disease, re-focusing therapeutic approaches by identifying the pernicious species of A beta ultimately responsible for the cognitive dysfunction that defines the disease.
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The 48 h total incubation leads to mature Aβ42 fibrils, as observed by TEM, while the Western blot shows the presence of higher order Aβ42 oligomers, likely due to an assembly-disassembly equilibrium that is established for mature Aβ fibrils.
“The 48 h total incubation leads to mature Aβ42 fibrils, as observed by TEM (Figure 10, panel a) while the Western blot shows the presence of higher order Aβ42 oligomers (Figure 10, lane a), likely due to an assembly-disassembly equilibrium that is established for mature Aβ fibrils.89–91”
From Bifunctional Compounds for Controlling Metal-Mediated Aggregation of the Aβ<sub>42</sub>Peptide (2012) · cites this paper as [91]
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Experimental — accuracy not yet measuredThe MOAB-2-based oAβ42 ELISA detects oligomeric but not monomeric Aβ42.
“In addition, the MOAB-2-based oA42 ELISA detects oligomeric but not monomeric A42 using synthetic preparations characterized previously by our laboratory (data not shown) (47,48).”
From APOE4-specific Changes in Aβ Accumulation in a New Transgenic Mouse Model of Alzheimer Disease (2012) · cites this paper as [48]
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“In addition, the MOAB-2-based oA 42 ELISA detects oligomeric but not monomeric A 42 using synthetic preparations characterized previously by our laboratory (data not shown) (47, 48).”
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Bharadwaj, P. R., Dubey, A. K., Masters, C. L., Martins, R. N., & Macreadie, I. G. (2008) · Journal of Cellular and Molecular Medicine · doi:10.1111/j.1582-4934.2009.00609.x · Cited by 147 · see citing works
Inhibiting, promoting, and preserving stability of functional proteinfibrils
Jones, O. G., & Mezzenga, R. (2011) · Soft Matter · doi:10.1039/c1sm06643a · Cited by 143 · see citing works
- Authors
- Stine, W. B., Dahlgren, K. N., Krafft, G. A., & LaDu, M. J.
- Affiliations
- Northwestern University (US)
- Corresponding author
- LaDu, Mary Jo
- ORCID
- LaDu, Mary Jo
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2003-03
- Volume
- 278
- Issue
- 13
- Pages
- 11612-11622
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2133631618
- Cited by
- 1,015
- References
- 74
- Citations by year
- 2026: 40 · 2025: 39 · 2024: 37 · 2023: 39 · 2022: 34 · 2021: 49 · 2020: 37 · 2019: 40 · 2018: 33 · 2017: 46 · 2016: 49 · 2015: 42 · 2014: 60 · 2013: 52 · 2012: 69
- FWCI
- 19.08
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Alzheimer's Association
- Northwestern University
- National Institutes of Health (1p01ag015501-01, 5r01ag013496-05)
- National Institute on Aging (R01AG019121, AG-15501, AG 19121, AG13496)
- Topics
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Health Sciences › Medicine › Psychiatry and Mental health › Dementia and Cognitive Impairment Research
- Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
- Keywords
- amyloid-beta, amyloid-beta 42, oligomerization, fibril formation, Alzheimer's disease, neurotoxicity, peptide aggregation, solution conditions, familial Alzheimer's disease mutations, amyloid plaques, peptide conformation
- MeSH terms
- Biopolymers, Circular Dichroism, Electrophoresis, Polyacrylamide Gel, Peptide Fragments, Solubility, Blotting, Western, Amyloid beta-Peptides
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