Amyloid Fibril Recognition with the Conformational B10 Antibody Fragment Depends on Electrostatic Interactions

Haupt, Christian; Morgado, Isabel; Kumar, Senthil T.; Parthier, C.; Bereza, Magdalena; Hortschansky, Peter; Stubbs, Milton T.; Horn, Uwe; Fändrich, Marcus

doi:10.1016/j.jmb.2010.10.059

Cited by 30 publications

(43 citation statements)

References 34 publications

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“…Subsequent research demonstrated that B10 recognizes amyloid fibrils through a pattern-recognition mechanism and binds to a strongly anionic surface moiety. This moiety is common to many different amyloid fibrils, enabling the poly-amyloid-specific binding of B10 to fibrils obtained from a broad range of different amino acid sequences (3,26).…”

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confidence: 99%

Molecular basis of β-amyloid oligomer recognition with a conformational antibody fragment

Morgado

Wieligmann

Bereza

et al. 2012

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

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103

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Oligomers are intermediates of the β-amyloid (Aβ) peptide fibrillogenic pathway and are putative pathogenic culprits in Alzheimer's disease (AD). Here we report the biotechnological generation and biochemical characterization of an oligomer-specific antibody fragment, KW1. KW1 not only discriminates between oligomers and other Aβ conformations, such as fibrils or disaggregated peptide; it also differentiates between different types of Aβ oligomers, such as those formed by Aβ (1-40) and Aβ (1-42) peptide. This high selectivity of binding contrasts sharply with many other conformational antibodies that interact with a large number of structurally analogous but sequentially different antigens. X-ray crystallography, NMR spectroscopy, and peptide array measurements imply that KW1 recognizes oligomers through a hydrophobic and significantly aromatic surface motif that includes Aβ residues 18-20. KW1-positive oligomers occur in human AD brain samples and induce synaptic dysfunctions in living brain tissues. Bivalent KW1 potently neutralizes this effect and interferes with Aβ assembly. By altering a specific step of the fibrillogenic cascade, it prevents the formation of mature Aβ fibrils and induces the accumulation of nonfibrillar aggregates. Our data illuminate significant mechanistic differences in oligomeric and fibril recognition and suggest the considerable potential of KW1 in future studies to detect or inhibit specific types of Aβ conformers.camelid antibody fragments | protein aggregation | immunotherapy

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confidence: 99%

Molecular basis of β-amyloid oligomer recognition with a conformational antibody fragment

Morgado

Wieligmann

Bereza

et al. 2012

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

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103

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“…The results of this systematic mutagenesis approach show that the basic residues (positively-charged residues) belonging to CDRs create a strongly positive electrostatic potential at the B10 binding surface, leading to electrostatic interactions with anionic groups present on some fibril surfaces. Moreover, the binding of B10 to amyloid fibrils composed of Ab 40 , insulin and G-helix peptide from sperm whale myoglobin is significantly reduced when the carboxyl groups of the fibrils are chemically modified [158]. The conformational specificity of B10 for amyloid fibrils seems therefore to depend upon specific electrostatic interactions with highly regular and anionic surface pattern common, at least locally, to different but not all amyloid fibrils, and not on the recognition of an amyloid generic backbone conformation.…”

Section: Generation Of Ab Specific Nanobodiesmentioning

confidence: 97%

“…From Refs. [155,158]. (CeD) Kinetics of fibril formation by Ab 40 monitored by ThT fluorescence and by TEM.…”

Section: Ab Peptide and Alzheimer's Diseasementioning

confidence: 99%

“…23 variants corresponding to single-site mutagenesis of the 23 residues within CDRs 1e3) to be produced and characterised, in order to understand the molecular basis of the specificity of B10 [158]. The results of this systematic mutagenesis approach show that the basic residues (positively-charged residues) belonging to CDRs create a strongly positive electrostatic potential at the B10 binding surface, leading to electrostatic interactions with anionic groups present on some fibril surfaces.…”

Section: Generation Of Ab Specific Nanobodiesmentioning

confidence: 99%

“…8B) [158]. The B10 molecular recognition reaction can therefore be described as an association between two relatively flat but complementary surfaces exhibiting opposite charges [158].…”

Section: Generation Of Ab Specific Nanobodiesmentioning

confidence: 99%

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Camelid single-domain antibody fragments: Uses and prospects to investigate protein misfolding and aggregation, and to treat diseases associated with these phenomena

2015

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Keywords:Variable domain of heavy-chain antibody Inhibition of protein misfolding and aggregation Protein misfolding diseases Amyloidoses V H H Nanobody a b s t r a c tThe deposition of misfolded peptides and proteins in the form of amyloid fibrils is the hallmark of nearly fifty medical disorders, including Alzheimer's disease, Parkinson's disease, prion diseases and type II diabetes. These disorders, referred to as amyloidoses, generally become apparent late in life. Their psycho-sociological and economic incidence in western societies will be therefore considerable in the coming decades due to the ageing of the population. Neither preventing nor curative treatments are available yet. These disorders constitute therefore a medical challenge of great importance. Thus, an extensive research is being carried out to understand, at the molecular level, (i) how amyloidogenic proteins misfold and convert from their soluble form into amyloid fibrils, and (ii) how these aggregates or some of their oligomeric precursor species are toxic. The formation of amyloid fibrils proceeds through a complex nucleation/polymerisation mechanism with the formation of various species, including small oligomers. In this review, we focus on how V H Hs or nanobodies, the antigen-binding domains of camelid heavy-chain antibodies, are being increasingly used to characterise each of the species formed on the pathway of fibril formation in terms of structure, stability, kinetics of formation and toxicity. We first introduce the characteristic features of nanobodies compared to those of conventional antibody fragments. Thereafter, we discuss how nanobodies, due to their unique properties, are used as probes to dissect the molecular mechanisms of misfolding and aggregation of six proteins associated with diseases, i.e. human lysozyme, b2-microglobulin, a-synuclein, prion, polyadenylate binding protein nuclear 1 and amyloid b-peptide. A brief general presentation of each disease and the associated peptide/protein is also provided. In addition, we discuss how nanobodies could be used as early diagnostic tools and as novel strategies to treat diseases associated with protein misfolding and aggregation.© 2015 Elsevier B.V. and Societe Française de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.Abbreviations: Ab, antibody; Ab, amyloid b-peptide; AD, Alzheimer's disease; ANS, anilino naphthalene-sulfonic acid; AP, alkaline phosphatase; APP, amyloid precursor protein; aSyn, a-synuclein; b2m, b2-microglobulin; BBB, bloodebrain barrier; CDR, complementarity determining region; C m , concentration of mid-denaturation; CR, Congo red; CSF, cerebrospinal fluid; DN6b2m, a truncated form of b2m lacking the six N-terminal amino acids; DLS, dynamic light scattering; DRA, dialysis-related amyloidosis; FR, framework; FTIR, Fourier transform infrared spectroscopy; Fv, variable fragment made of the VH and VL domains of conventional antibodies; GFP, green fluorescent protein; HCAb, heavy-chain antibody; H/D, hydrogen/deuterium; HSQC, heteronuclear s...

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Amyloid‐Like Fibrils: Origin, Structure, Properties, and Potential Technological Applications

Taboada

Barbosa

Juárez

et al. 2013

Proteins in Solution and at Interfaces

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Amyloid Fibril Recognition with the Conformational B10 Antibody Fragment Depends on Electrostatic Interactions

Cited by 30 publications

References 34 publications

Molecular basis of β-amyloid oligomer recognition with a conformational antibody fragment

Molecular basis of β-amyloid oligomer recognition with a conformational antibody fragment

Camelid single-domain antibody fragments: Uses and prospects to investigate protein misfolding and aggregation, and to treat diseases associated with these phenomena

Amyloid‐Like Fibrils: Origin, Structure, Properties, and Potential Technological Applications

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