Structural studies of amyloid-β peptides: Unlocking the mechanism of aggregation and the associated toxicity

Aleksis, Rihards; Oleskovs, Filips; Jaudzems, Kristaps; Pahnke, Jens; Biverstål, Henrik

doi:10.1016/j.biochi.2017.07.011

Cited by 61 publications

(56 citation statements)

References 166 publications

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“…This is probably one important reason behind the fact that, despite immense efforts, the exact mechanisms of Aβ self-assembly are still unknown 9-11 . Recent advances have however revealed new insights into the nucleation mechanism of Aβ in vitro 12,13 , structural details of the fibril morphology 14 and biological mechanisms implicated in the AD etiopathology 15,16 . These experiments typically require access to very pure and homogeneous Aβ peptides as small impurities or preformed seeds have a great impact on the aggregation behavior 17 .…”

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High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain

Abelein

Chen

Aleksis

et al. 2020

Sci Rep

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During storage in the silk gland, the n-terminal domain (nt) of spider silk proteins (spidroins) keeps the aggregation-prone repetitive region in solution at extreme concentrations. We observe that nts from different spidroins have co-evolved with their respective repeat region, and now use an NT that is distantly related to previously used NTs, for efficient recombinant production of the amyloid-β peptide (Aβ) implicated in Alzheimer's disease. A designed variant of nt from Nephila clavipes flagelliform spidroin, which in nature allows production and storage of β-hairpin repeat segments, gives exceptionally high yields of different human Aβ variants as a solubility tag. This tool enables efficient production of target peptides also in minimal medium and gives up to 10 times more isotope-labeled monomeric Aβ peptides per liter bacterial culture than previously reported.Orb-weaving spiders manufacture up to seven different silks, e.g. dragline silk derived from major ampullate silk proteins (spidroins, MaSp) and flagelliform silk derived from flagelliform spidroins (FlSp). The various spidroins share a common architecture -a large core repetitive region capped by globular N-and C-terminal domains (NT and CT) 1 . The divergent and large aggregation-prone repetitive regions of the spidroins determine the mechanical properties of the respective spider silks, while the terminal domains regulate silk fiber formation 2,3 . Despite their high aggregation propensity the spidroins can be stored at extremely high concentrations (30-50% w/v) in the spider silk gland, solubilized by the NT domain 1,4 .The NT dimerizes upon a drop in pH, which is crucial for silk fiber formation 1,5 . To ensure solubility also at low pH and widen the applicability of NT as a solubility enhancing fusion partner, a charged-reversed mutant has been designed (referred to as NT* MaSp ) 6 . The previously reported NT* MaSp tag is derived from the NT domain of Euprosthenops australis MaSp1 and folds as a five-helix bundle 6,7 . NT* MaSp is a pH insensitive constitutive monomer, highly stable and extremely soluble, and has been successfully applied for efficient production and purification of, among others, lung surfactant protein analogs, cholecystokinin-58, human antimicrobial cathelicidin and a designed β-sheet protein 6,8 .Aggregation-prone proteins and peptides are associated with several neurodegenerative disorders, e.g. Alzheimer's disease (AD), the most prevalent form of dementia 9,10 . These proteins/peptides often exhibit high β-sheet propensity, which make them prone to aggregate and form insoluble amyloid fibrils 11 . These intrinsic properties of amyloid-forming proteins make high-yield biochemical production challenging, yet the availability of pure protein samples is crucial for studying protein self-assembly and its associated neurotoxicity in vitro and in vivo. This is probably one important reason behind the fact that, despite immense efforts, the exact mechanisms of Aβ self-assembly are still unknown 9-11 . Recent advances have howe...

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confidence: 99%

High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain

Abelein

Chen

Aleksis

et al. 2020

Sci Rep

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“…The mechanism of how these structures arise, diversify, and propagate remains under active investigation. 5−8 Ostwald’s rule of stages suggests that a less stable phase may appear first and accelerate the formation of more stable phases. 9,10 Evidence obtained with simple peptides suggests that initial liquid–liquid transitions create metastable phases where peptides access β-sheets.…”

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Multistep Conformation Selection in Amyloid Assembly

et al. 2017

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“…The most widely accepted hypothesis to explain the pathogenesis of Alzheimer's disease (AD) is the amyloid cascade, formalized by Hardy and Higgins , which states that β‐sheeted Aβ proteins aggregate and deposit as extracellular plaques generated by the proteolytic cleavage of the amyloid precursor protein (APP) and intracellular tangles as central drivers of disease progression . Early‐onset AD induced by Aβ mutations provide a strong link between pathogenesis and protein aggregation .…”

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confidence: 99%

“…Alzheimer's disease (AD) is the most prevalent age‐related cause of dementia representing about 50 % to 70 % of the total cases . Because the primary risk factor for AD is age, the prevalence of the disease is increasing dramatically with the ageing populations worldwide .…”

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“…Subsequent tau aggregation mediates synaptic dysfunction and neuronal death, which are the molecular signatures of underlying cognitive decline . However, several lines of evidence indicate that not insoluble precipitation of larger aggregates such as senile plaques or intracellular neurofibrillary tangles rather than small soluble and highly neurotoxic Aβ oligomers are the key drivers for AD pathogenesis .…”

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Advancements of the sFIDA method for oligomer‐based diagnostics of neurodegenerative diseases

et al. 2018

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Early diagnosis of Alzheimer's disease (AD) is of great importance for the development of therapeutics and their application in the clinical environment. Amyloid β (Aβ) oligomers are crucial for the onset and progression of AD and represent a popular drug target, being presumably the most direct biomarker. Efforts to measure Aβ oligomers in body fluids are hampered by the low analyte concentration and presence of Aβ monomers. The surface‐based fluorescence intensity distribution analysis (sFIDA) features both highly specific and sensitive oligomer quantitation as well as total insensitivity towards monomers. In this Review, we highlight structural features of oligomeric and fibrillar Aβ. Recent advancements in sFIDA assay development have been the successful automation, adaption for additional biomarkers such as α‐synuclein oligomers, and significant improvement of essential assay parameters.

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Structural studies of amyloid-β peptides: Unlocking the mechanism of aggregation and the associated toxicity

Cited by 61 publications

References 166 publications

High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain

High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain

Multistep Conformation Selection in Amyloid Assembly

Advancements of the sFIDA method for oligomer‐based diagnostics of neurodegenerative diseases

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