Selection and characterization of Affibody ligands binding to Alzheimer amyloid β peptides

Grönwall, Caroline; Jonsson, Andreas; Lindström, Sara; Gunneriusson, Elin; Ståhl, Stefan; Herne, Nina

doi:10.1016/j.jbiotec.2006.09.013

Cited by 111 publications

(123 citation statements)

References 62 publications

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“…This has been observed in other SPR experiments on Aβ40 interactions. 45 For each protein, one low concentration with significant binding was selected for curve fitting ( Figure 6) to avoid the effects of a second weaker binding process (see Figure S5). The dissociation, apart from the initial step, is very slow for all three proteins.…”

Section: ■ Resultsmentioning

confidence: 99%

Charge Dependent Retardation of Amyloid β Aggregation by Hydrophilic Proteins

Assarsson

Hellstrand

Cabaleiro-Lago

et al. 2014

ACS Chem. Neurosci.

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The aggregation of amyloid β peptides (Aβ) into amyloid fibrils is implicated in the pathology of Alzheimer's disease. In light of the increasing number of proteins reported to retard Aβ fibril formation, we investigated the influence of small hydrophilic model proteins of different charge on Aβ aggregation kinetics and their interaction with Aβ. We followed the amyloid fibril formation of Aβ40 and Aβ42 using thioflavin T fluorescence in the presence of six charge variants of calbindin D 9k and singlechain monellin. The formation of fibrils was verified with transmission electron microscopy. We observe retardation of the aggregation process from proteins with net charge +8, +2, −2, and −4, whereas no effect is observed for proteins with net charge of −6 and −8. The single-chain monellin mutant with the highest net charge, scMN+8, has the largest retarding effect on the amyloid fibril formation process, which is noticeably delayed at as low as a 0.01:1 scMN+8 to Aβ40 molar ratio. scMN+8 is also the mutant with the fastest association to Aβ40 as detected by surface plasmon resonance, although all retarding variants of calbindin D 9k and single-chain monellin bind to Aβ40.

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Section: ■ Resultsmentioning

confidence: 99%

Charge Dependent Retardation of Amyloid β Aggregation by Hydrophilic Proteins

Assarsson

Hellstrand

Cabaleiro-Lago

et al. 2014

ACS Chem. Neurosci.

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“…The hairpin is predicted to comprise residues 17 to 23 and 30 to 36 as antiparallel β-strands connected by a turn involving residues 25 to 29. There is considerable evidence that such a conformation is accessible in monomeric Aβ: (i) it forms in complex with binding proteins (16,17), (ii) its secondary structure elements persistently appear in computer simulations of different Aβ fragments (15,24,25), and (iii) NMR data suggest that turn formation of residues 24 to 28 nucleates monomer folding (26). We previously postulated that metastable Aβ oligomers contain hairpin subunits and that conversion into a related cross-β conformation transforms oligomers into fibril seeds and primes these for the runaway aggregation that is typical of amyloid fibril formation.…”

Section: Discussionmentioning

confidence: 99%

“…Such a "hairpin" conformation (Fig. 1A) is in fact also induced when Aβ forms a complex with a phage-display selected Affibody-binding protein (16,17). The hairpin is topologically similar to the conformation of Aβ in fibrils.…”

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confidence: 95%

Stabilization of neurotoxic Alzheimer amyloid-β oligomers by protein engineering

Sandberg

Luheshi²,

Söllvander³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

323

405

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Soluble oligomeric aggregates of the amyloid-β peptide (Aβ) have been implicated in the pathogenesis of Alzheimer's disease (AD). Although the conformation adopted by Aβ within these aggregates is not known, a β-hairpin conformation is known to be accessible to monomeric Aβ. Here we show that this β-hairpin is a building block of toxic Aβ oligomers by engineering a doublecysteine mutant (called AβCC) in which the β-hairpin is stabilized by an intramolecular disulfide bond. Aβ 40 CC and Aβ 42 CC both spontaneously form stable oligomeric species with distinct molecular weights and secondary-structure content, but both are unable to convert into amyloid fibrils. Biochemical and biophysical experiments and assays with conformation-specific antibodies used to detect Aβ aggregates in vivo indicate that the wild-type oligomer structure is preserved and stabilized in AβCC oligomers. Stable oligomers are expected to become highly toxic and, accordingly, we find that β-sheet-containing Aβ 42 CC oligomers or protofibrillar species formed by these oligomers are 50 times more potent inducers of neuronal apoptosis than amyloid fibrils or samples of monomeric wild-type Aβ 42 , in which toxic aggregates are only transiently formed. The possibility of obtaining completely stable and physiologically relevant neurotoxic Aβ oligomer preparations will facilitate studies of their structure and role in the pathogenesis of AD. For example, here we show how kinetic partitioning into different aggregation pathways can explain why Aβ 42 is more toxic than the shorter Aβ 40 , and why certain inherited mutations are linked to protofibril formation and early-onset AD.amyloid-β peptide | protein aggregation | protein structure | protofibril | β-hairpin conformation

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“…An interesting case is an Affibody molecule specific for the amyloid-b (Ab) peptide involved in Alzheimer's disease [72]. According to the amyloid hypothesis, the pathogenesis of Alzheimer's disease is associated with the oligomerization and aggregation of the Ab peptide into protein plaques.…”

Section: Blocking Applicationsmentioning

confidence: 99%

Affibody molecules: Engineered proteins for therapeutic, diagnostic and biotechnological applications

et al. 2010

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a b s t r a c tAffibody molecules are a class of engineered affinity proteins with proven potential for therapeutic, diagnostic and biotechnological applications. Affibody molecules are small (6.5 kDa) single domain proteins that can be isolated for high affinity and specificity to any given protein target. Fifteen years after its discovery, the Affibody technology is gaining use in many groups as a tool for creating molecular specificity wherever a small, engineering compatible tool is warranted. Here we summarize recent results using this technology, propose an Affibody nomenclature and give an overview of different HER2-specific Affibody molecules. Cumulative evidence suggests that the three helical scaffold domain used as basis for these molecules is highly suited to create a molecular affinity handle for vastly different applications.

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Selection and characterization of Affibody ligands binding to Alzheimer amyloid β peptides

Cited by 111 publications

References 62 publications

Charge Dependent Retardation of Amyloid β Aggregation by Hydrophilic Proteins

Charge Dependent Retardation of Amyloid β Aggregation by Hydrophilic Proteins

Stabilization of neurotoxic Alzheimer amyloid-β oligomers by protein engineering

Affibody molecules: Engineered proteins for therapeutic, diagnostic and biotechnological applications

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