A kinetic study on the aggregation behavior of β-amyloid peptides in different initial solvent environments

Amyloid beta peptide (Abeta) fibril formation is widely believed to be the causative event of Alzheimer's disease pathogenesis. Therapeutic approaches are therefore in development that target various sites in the production and aggregation of Abeta. Herein we present a high-throughput screening tool to generate novel hit compounds that block Abeta fibril formation. This tool is an application for our fibril model (Abeta(16-37)Y(20)K(22)K(24))(4), which is a covalent assembly of four Abeta fragments. With this tool, screening studies are complete within one hour, as opposed to days with native Abeta(1-40). A Z' factor of 0.84+/-0.03 was determined for fibril formation and inhibition, followed by the reporter molecule thioflavin T. Herein we also describe the analysis of a broad range of reported inhibitors and non-inhibitors of Abeta fibril formation to test the validity of the system.

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“…[52] Measurement of (Ab . The concentrations of studied inhibitors were 0, 0.01, 0.1, 1, 10, and 100 mm.…”

Section: Measurement Of Ab 1-40 Aggregationmentioning

confidence: 99%

Designed Amyloid β Peptide Fibril—A Tool for High‐Throughput Screening of Fibril Inhibitors

et al. 2007

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“…Mounting evidence originating from in vitro toxicity studies with synthetic Aβ peptides shows that Aβ, in an aggregated state (fi bril, protofi bril, low molecular-weight oligomer, or diffusible, non-fi brillar ligand), is toxic to neurons in cultures (Bieschke et al 2008;Stefani 2007;Baglioni et al 2006;Dobson 2006). The assembly phenomenon is reported to be dependent on the pH of solutions, concentration of Aβs and incubation time in solution (Wei and Shea 2006;Wang et al 2004). Several β-amyloid fragments have been reported to form such assemblies, but only Aβs that include a substantial portion of the transmembrane sequence were showed to be able to assemble into aggregates that are stable at pH 7.4 and resistant to disruption by sodium dodecylsulphate (SDS) (Burdick et al 1992;Pike et al 1993Pike et al , 1995.…”

Section: Introductionmentioning

confidence: 99%

Rapid aggregation and assembly in aqueous solution of Aβ (25–35) peptide

et al. 2009

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The highly toxic A beta (25-35) is a peculiar peptide that differs from all the other commonly studied beta-amyloid peptides because of its extremely rapid aggregation properties and enhanced neurotoxicity. We investigated A beta (25-35) aggregation in H2O at pH 3.0 and at pH 7.4 by means of in-solution analyses. Adopting UV spectroscopy, Congo red spectrophotometry and thioflavin T fluorimetry, we were able to quantify, in water, the very fast assembling time necessary for A beta (25-35) to form stable insoluble aggregates and their ability to seed or not seed fibril growth. Our quantitative results, which confirm a very rapid assembly leading to stable insoluble aggregates of A beta (25-35) only when incubated at pH 7.4, might be helpful for designing novel aggregation inhibitors and to shed light on the in vivo environment in which fibril formation takes place.

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“…As the bounded molecules accumulate at these sites, the size of the protein assemblies gradually increases [24,25]. Evidence showed that the mechanism of protein aggregation is described as a process of nucleation followed by elongation and growth [26][27][28][29]. Results from past studies showed that the aggregation behavior of proteins can be altered by a number of factors including: (i) external/environmental factors: protein concentration, temperature, presence of salt, metal ion concentration, type of dissolving solvent, and pH of solution and (ii) internal/structural factors: hydrophobicity, β-sheet secondary structure content [10,20,30,31].…”

Section: Cause Characteristics and Detection Of Protein Aggregatesmentioning

confidence: 99%

“…Whether in water or physiological buffers, under higher concentrations and/or high temperature, reversible aggregation into higher molecular weight species may occur [95,98]. Moreover, the effect of dissolving solvents on the kinetic behavior of Aβ aggregation was quantitatively analyzed [26]. It was found that the morphological feature of fibrils/aggregates formed was also dependent on the solvent type (see Fig.…”

Section: Aβ Aggregation Processmentioning

confidence: 99%

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Diseases of protein aggregation and the hunt for potential pharmacological agents

Wang

Yamamoto

et al. 2008

Biotechnology Journal

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Protein aggregation is a ubiquitous phenomenon significant to all aspects of science. Notably, the formation of protein aggregates is frequently encountered in biochemical research and biopharmaceutical industry. Formation of protein aggregates is generally regarded to be associated with partially folded intermediate species that are susceptible to self-association due to the exposure of hydrophobic core. Evidence supports the concept that the formation of aggregates in vitro is a generic property of proteins. In human etiology, more than 20 different devastating human diseases have been reported to be associated with protein aggregation. Although protein aggregation diseases have been the center of intense research, much remains to be learned regarding the underlying molecular mechanisms. In this review, the general background information on protein aggregation is first provided. Next, we summarize the properties, characteristics and causes of protein aggregates. Finally, from the perspectives of epidemiology, pathogenesis, existing mechanisms, relevant hypotheses, and current as well as potential therapeutic approaches, two examples of protein aggregation diseases, Alzheimer's disease and cataract, are briefly discussed. Importantly, while a variety of molecules have been suggested, the effective therapeutic drugs for curing the diseases involving protein aggregation have yet to be identified. We believe that a better understanding of the mechanisms of protein aggregation process and an extensive investigation into the drug penetration, efficacy, and side effects will certainly aid in developing the successful pharmacological agents for these diseases.

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A kinetic study on the aggregation behavior of β-amyloid peptides in different initial solvent environments

Cited by 20 publications

References 52 publications

Designed Amyloid β Peptide Fibril—A Tool for High‐Throughput Screening of Fibril Inhibitors

Designed Amyloid β Peptide Fibril—A Tool for High‐Throughput Screening of Fibril Inhibitors

Rapid aggregation and assembly in aqueous solution of Aβ (25–35) peptide

Diseases of protein aggregation and the hunt for potential pharmacological agents

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