Reduced Global Cooperativity is a Common Feature Underlying the Amyloidogenicity of Pathogenic Lysozyme Mutations

Dumoulin, Mireille; Canet, Denis; Last, Alexander M.; Pardon, Els; Archer, David B.; Wyns, Lode; Matagne, André; Robinson, Carol V.; Redfield, Christina; Dobson, Christopher M.

doi:10.1016/j.jmb.2004.11.020

Cited by 105 publications

(212 citation statements)

References 35 publications

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“…These latter peaks are not observed in the mass spectra of the D67H variant in the presence of cAb-HuL6, indicating that the binding of this nanobody restores the global cooperativity of the D67H variant. The same inhibition was observed for cAb-HuL6 and the I56T variant [82], and for cAb-HuL22 and both the I56T and D67H variants [44]. On the other hand, these yellow peaks are observed in the mass spectra of the D67H variant in the presence of cAb-HuL5; the binding of cAb-HuL5 does therefore not inhibit the was added between the two b-sheets thanks to the S54C and I78C mutations (IMGT numbering) [70,71].…”

Section: Human Lysozyme and Systemic Amyloidosissupporting

confidence: 54%

“…3D). In this species, the b-domain and the C-helix (referred to as the amylotope [44]) are cooperatively unfolded while the rest of the adomain remains native [81,82] (Fig. 3A).…”

Section: Human Lysozyme and Systemic Amyloidosismentioning

confidence: 99%

“…Indeed, the amyloidogenic lysozyme variants must be incubated under rather harsh denaturing conditions (i.e. in 0.1 M citrate buffer pH 5.5 containing 3 M urea at 48 C or in 0.1 M acetate buffer pH 5.0 at 65 C) to form amyloid fibrils in vitro in a reasonable time [69,82]; thus, in order to use antibody fragments as molecular structural and mechanistic probes they must be stable and functional under those conditions.…”

Section: Human Lysozyme and Systemic Amyloidosismentioning

confidence: 99%

“…0.1 M citrate buffer pH 5.5 containing 3 M urea at 48 C) [69,82,83] (Table 1). On the contrary, cAb-HuL22 is less thermostable than the lysozyme variants and could therefore not be used to inhibit fibril formation [44].…”

Section: Human Lysozyme and Systemic Amyloidosismentioning

confidence: 99%

“…In the presence of an equimolar amount of cAb-HuL6, the D67H (Fig. 3C) and I56T [69,82] variants of lysozyme do not significantly aggregate, even after 24 h of incubation, while both variants completely aggregate into amyloid fibrils in less than 5 h when incubated in the absence of nanobodies [69,82]. The binding of cAb-HuL22-S54C/I78C and cAbHuL5G to D67H also significantly inhibits fibril formation although, due to their low affinity under the conditions used to trigger lysozyme fibril formation, a high nanobody/lysozyme ratio (i.e.…”

Section: Cab-hul5mentioning

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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Section: Human Lysozyme and Systemic Amyloidosissupporting

confidence: 54%

“…3D). In this species, the b-domain and the C-helix (referred to as the amylotope [44]) are cooperatively unfolded while the rest of the adomain remains native [81,82] (Fig. 3A).…”

Section: Human Lysozyme and Systemic Amyloidosismentioning

confidence: 99%

Section: Human Lysozyme and Systemic Amyloidosismentioning

confidence: 99%