Early Kinetics of Amyloid Fibril Formation Reveals Conformational Reorganisation of Initial Aggregates

Cerdà-Costa, Núria; Esteras-Chopo, Alexandra; Avilés, Francesc X.; Serrano, Luís; Villegas, Sandra

doi:10.1016/j.jmb.2006.12.007

Cited by 60 publications

(74 citation statements)

References 53 publications

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“…The NDP kinases have in addition the opposite side of the beta sheet covered by the helix α4 which is not protected from proteinase K digestion in the aggregated conformation. The β-strand predicted to be most amyloidogenic has a similar position in NDPK-HA and in the abovementioned proteins (Cerda-Costa et al 2007). …”

Section: Discussionmentioning

confidence: 91%

Aggregation of the neuroblastoma-associated mutant (S120G) of the human nucleoside diphosphate kinase-A/NM23-H1 into amyloid fibrils

Georgescauld

Sabaté

Espargaró

et al. 2011

Naunyn-Schmiedeberg's Arch Pharmacol

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The human nucleoside diphosphate (NDP) kinase A, product of the NME1 gene also named NM23-H1, is known as a metastasis suppressor protein. A naturally occurring variant, S120G, identified in neuroblastomas, possesses native three-dimensional structure and enzymatic activity but displays reduced conformational stability and a folding defect with the accumulation of a "molten globule" folding intermediate during refolding in vitro. As such intermediate has been postulated to be involved in amyloid formation, NDP kinase A may serve as a model protein for studying the relationship between folding intermediates and amyloid fibrils. The NDP kinase A S120G was heated in phosphate buffer (pH 7.0). The protein precipitated as amyloid fibrils, as demonstrated by electron microscopy, Congo red, and thioflavin T binding and FTIR spectroscopy. The NDP kinase A S120G, at neutral pH and at moderate temperature experiences a transition towards amyloid fibrils. The aggregation process was faster if seeded by preformed fibrils. The fibrils presented a large proteinase K-resistant core not including residue Gly 120, as shown by mass spectrometry. This suggests that the aggregation process is triggered by the reduced stability of the S120G variant and not by a specific increase in the kinase domain intrinsic aggregation propensity at the place of mutation. This constitutes one of the few reports on a protein involved in cancer biology able to aggregate into amyloid structures under mild conditions.

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Section: Discussionmentioning

confidence: 91%

Aggregation of the neuroblastoma-associated mutant (S120G) of the human nucleoside diphosphate kinase-A/NM23-H1 into amyloid fibrils

Georgescauld

Sabaté

Espargaró

et al. 2011

Naunyn-Schmiedeberg's Arch Pharmacol

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“…Previous studies indicate that β-sheet formation is a crucial early step in the amyloidogenesis of α-syn 38,39 and is a signature of amyloid fibril formation. To evaluate the effects of Sel on β-sheet formation, we performed circular dichroism (CD) measurements (Fig.…”

Section: Resultsmentioning

confidence: 99%

The Anti-Parkinsonian Drug Selegiline Delays the Nucleation Phase of α-Synuclein Aggregation Leading to the Formation of Nontoxic Species

Braga

Follmer

Palhano

et al. 2011

Journal of Molecular Biology

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Parkinson's disease (PD) is a movement disorder characterized by the loss of dopaminergic neurons in the substantia nigra and the formation of intraneuronal inclusions called Lewy bodies, which are composed mainly of α-synuclein (α-syn). Selegiline (Sel) is a noncompetitive monoamino oxidase B inhibitor that has neuroprotective effects and has been administered to PD patients as monotherapy or in combination with L-dopa. Besides its known effect of increasing the level of dopamine (DA) by monoamino oxidase B inhibition, Sel induces other effects that contribute to its action against PD. We evaluated the effects of Sel on the in vitro aggregation of A30P and wild-type α-syn. Sel delays fibril formation by extending the lag phase of aggregation. In the presence of Sel, electron microscopy reveals amorphous heterogeneous aggregates, including large annular species, which are innocuous to a primary culture enriched in dopaminergic neurons, while their age-matched counterparts are toxic. The inhibitory effect displayed by Sel is abolished when seeds (small fibril pieces) are added to the aggregation reaction, reinforcing the hypothesis that Sel interferes with early nuclei formation and, to a lesser extent, with fibril elongation. NMR experiments indicate that Sel does not interact with monomeric α-syn. Interestingly, when added in combination with DA (which favors the formation of toxic protofibrils), Sel overrides the *Corresponding author. E-mail address: foguel@bioqmed.ufrj.br.† C.A.B. and C.F. contributed equally to this work. Present address: C. Follmer, Instituto de Química, Departamento de Físico-Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.Abbreviations used: PD, Parkinson's disease; a-syn, a-synuclein; Sel, selegiline; DA, dopamine; MAO-B, monoamino oxidase B; MPTP, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; ROS, reactive oxygen species; DA, dopamine; WT, wild type; Thio-T, thioflavin T; TTR, transthyretin; EM, electron microscopy; HSQC, heteronuclear single quantum coherence; A30P-F, preformed A30P fibrils; LS, light scattering; DAPI, 4V, 6-diamidino-2-phenylindole; EGCG, epigallocatechin gallate; PBS, phosphate-buffered saline. inhibitory effect of DA and favors fibrillation. Additionally, Sel blocks the formation of smaller toxic aggregates by perturbing DA-dependent fibril disaggregation. These effects might be beneficial for PD patients, since the sequestration of protofibrils into fibrils or the inhibition of fibril dissociation could alleviate the toxic effects of protofibrils on dopaminergic neurons. In nondopaminergic neurons, Sel might slow the fibrillation, giving rise to the formation of large nontoxic aggregates.

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“…35,[43][44][45] Putatively, the structural rearrangement stage is important in the formation of strong inter-protein b-sheet contacts that help to stabilize aggregates and make them effectively irreversible. 13,46,47 This is arguably the least well understood stage of non-native aggregation, due in large part to experimental and theoretical difficulties with isolating or directly monitoring U x or A x . [43][44][45]48 Therefore, for simplicity the step U x !…”

Section: Aggregation Kinetics and Shelf Lifementioning

confidence: 99%

Principles, approaches, and challenges for predicting protein aggregation rates and shelf life

Weiss

Young

Roberts

2009

Journal of Pharmaceutical Sciences

275

289

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Early Kinetics of Amyloid Fibril Formation Reveals Conformational Reorganisation of Initial Aggregates

Cited by 60 publications

References 53 publications

Aggregation of the neuroblastoma-associated mutant (S120G) of the human nucleoside diphosphate kinase-A/NM23-H1 into amyloid fibrils

Aggregation of the neuroblastoma-associated mutant (S120G) of the human nucleoside diphosphate kinase-A/NM23-H1 into amyloid fibrils

The Anti-Parkinsonian Drug Selegiline Delays the Nucleation Phase of α-Synuclein Aggregation Leading to the Formation of Nontoxic Species

Principles, approaches, and challenges for predicting protein aggregation rates and shelf life

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