Unraveling amyloid toxicity pathway in NIH3T3 cells by a combined proteomic and<sup>1</sup>H‐NMR metabonomic approach

Vilasi, Annalisa; Vilasi, Silvia; Romano, R.; Acernese, F.; Barone, F.; Balestrieri, Maria Luisa; Maritato, Rosa; Irace, Gaetano; Sirangelo, Ivana

doi:10.1002/jcp.24294

Cited by 10 publications

(16 citation statements)

References 72 publications

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“…As expected, in the absence of D-ribose, early prefibrillar aggregates (0 and 5 days) strongly affected cell viability (about 40% MTT reduction), whereas fibrillar aggregates (10 days) did not ( Figure 8 ) in accordance with previous studies [20], [22]. Conversely, aggregates formed in the presence of D-ribose after 5 days of incubation did not affect cell viability supporting our finding that D-ribose induces rapid formation of harmless fibrillar aggregates.…”

Section: Resultssupporting

confidence: 92%

“…In particular, the replacement of both indole residues located at positions 7 and 14 in the N-terminal region with phenylalanine (W7FW14F) renders myoglobin highly susceptible to aggregation and able to form amyloid fibrils under physiological conditions of pH and temperature [16]–[19]. Similarly to other amyloidogenic proteins, the cytotoxicity of the W7FW14F apomyoglobin aggregates is associated to the early oligomers intermediates rather than mature fibrils [20]–[22]. These features make the W7FW14F myoglobin a suitable model for studying the molecular mechanisms underlying amyloid aggregation under physiological conditions.…”

Section: Introductionmentioning

confidence: 99%

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Glycation Accelerates Fibrillization of the Amyloidogenic W7FW14F Apomyoglobin

et al. 2013

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Neurodegenerative diseases are associated with misfolding and deposition of specific proteins, either intra or extracellularly in the nervous system. Advanced glycation end products (AGEs) originate from different molecular species that become glycated after exposure to sugars. Several proteins implicated in neurodegenerative diseases have been found to be glycated in vivo and the extent of glycation is related to the pathologies of the patients. Although it is now accepted that there is a direct correlation between AGEs formation and the development of neurodegenerative diseases, several questions still remain unanswered: whether glycation is the triggering event or just an additional factor acting on the aggregation pathway. To this concern, in the present study we have investigated the effect of glycation on the aggregation pathway of the amyloidogenic W7FW14F apomyoglobin. Although this protein has not been related to any amyloid disease, it represents a good model to resemble proteins that intrinsically evolve toward the formation of amyloid aggregates in physiological conditions. We show that D-ribose, but not D-glucose, rapidly induces the W7FW14F apomyoglobin to generate AGEs in a time-dependent manner and protein ribosylation is likely to involve lysine residues on the polypeptide chain. Ribosylation of the W7FW14F apomyoglobin strongly affects its aggregation kinetics producing amyloid fibrils within few days. Cytotoxicity of the glycated aggregates has also been tested using a cell viability assay. We propose that ribosylation in the W7FW14F apomyoglobin induces the formation of a cross-link that strongly reduces the flexibility of the H helix and/or induce a conformational change that favor fibril formation. These results open new perspectives for AGEs biological role as they can be considered not only a triggering factor in amyloidosis but also a player in later stages of the aggregation process.

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Section: Resultssupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Glycation Accelerates Fibrillization of the Amyloidogenic W7FW14F Apomyoglobin

et al. 2013

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“…We demonstrated that the cytotoxicity induced by W7FW14F ApoMb is closely related to oxidative stress [Sirangelo et al, ; Vilasi et al, ]. Similarly to previous studies aimed at evaluating the mechanisms of amyloid‐β‐induced neurotoxicity [Bate et al, ; Li et al, ], our data show that the time‐dependent decrease of cell viability elicited by W7FW14F ApoMb insult is prevented by PAF‐R antagonist, CV3988, thus suggesting the involvement of PAF signaling in the apoptotic pathway activated by W7FW14F ApoMb.…”

Section: Discussionsupporting

confidence: 84%

“…We first evaluated the oxidative stress induced by W7FW14F ApoMb amyloid oligomers, known to occur at early time of cell exposure to aggregates [Sirangelo et al, ; Vilasi et al, ]. Results indicated that exposure of SH‐SY5Y cells to W7FW14F ApoMb aggregates up to 360 min led to a significant increase of intracellular ROS compared with untreated cells ( P < 0.05 at 15, 30, and 60 min, P < 0.01 at 180 and 360 min) (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The cytotoxicity induced by W7FW14F ApoMb amyloid oligomers is reported to be closely related to oxidative stress, as well as perturbation of calcium homeostasis, apoptotic and survival pathways activation, and membrane damage [Sirangelo et al, 2009;Vilasi et al, 2013]. In this work, we examined the possible involvement of PAF signaling pathway in the cytotoxicity induced by W7FW14F ApoMb amyloid aggregates.…”

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Platelet‐Activating Factor Mediates the Cytotoxicity Induced by W7FW14F Apomyoglobin Amyloid Aggregates in Neuroblastoma Cells

Sirangelo

Giovane

Maritato

et al. 2014

J of Cellular Biochemistry

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W7FW14F apomyoglobin (W7FW14F ApoMb) amyloid aggregates induce cytotoxicity in SH-SY5Y human neuroblastoma cells through a mechanism not fully elucidated. Amyloid neurotoxicity process involves calcium dyshomeostasis and reactive oxygen species (ROS) production. Another key mediator of the amyloid neurotoxicity is Platelet-Activating Factor (PAF), an inflammatory phospholipid implicated in neurodegenerative diseases. Here, with the aim at evaluating the possible involvement of PAF signaling in the W7FW14F ApoMb-induced cytotoxicity, we show that the presence of CV3899, a PAF receptor (PAF-R) antagonist, prevented the detrimental effect of W7FW14F ApoMb aggregates on SH-SY5Y cell viability. Noticeably, we found that the activation of PAF signaling, following treatment with W7FW14F ApoMb, involves a decreased expression of the PAF acetylhydroase II (PAF-AH II). Interestingly, the reduced PAF-AH II expression was associated with a decreased acetylhydrolase (AH) activity and to an increased sphingosine-transacetylase activity (TA(S)) with production of N-acetylsphingosine (C2-ceramide), a well known mediator of neuronal caspase-dependent apoptosis. These findings suggest that an altered PAF catabolism takes part to the molecular events leading to W7FW14F ApoMb amyloid aggregates-induced cell death.

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Glycation of Wild‐Type Apomyoglobin Induces Formation of Highly Cytotoxic Oligomeric Species

Iannuzzi

Carafa

Altucci

et al. 2015

Journal Cellular Physiology

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Protein glycation is a non-enzymatic, irreversible modification of protein amino groups by reactive carbonyl species leading to the formation of advanced glycation end products (AGEs). Several proteins implicated in neurodegenerative diseases have been found to be glycated in vivo and the extent of glycation is related to the pathologies of the patients. Although it is now accepted that there is a direct correlation between AGEs formation and the development of neurodegenerative diseases related to protein misfolding and amyloid aggregation, several questions still remain unanswered: whether glycation is the triggering event or just an additional factor acting on the aggregation pathway. We have recently shown that glycation of the amyloidogenic W7FW14F apomyoglobin mutant significantly accelerates the amyloid fibrils formation providing evidence that glycation actively participates to the process. In the present study, to test if glycation can be considered also a triggering factor in amyloidosis, we evaluated the ability of different glycation agents to induce amyloid aggregation in the soluble wild-type apomyoglobin. Our results show that glycation covalently modifies apomyoglobin and induces conformational changes that lead to the formation of oligomeric species that are not implicated in amyloid aggregation. Thus, AGEs formation does not trigger amyloid aggregation in the wild-type apomyoglobin but only induce the formation of soluble oligomeric species able to affect cell viability. The molecular bases of cell toxicity induced by AGEs formed upon glycation of wild-type apomyoglobin have been also investigated.

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Unraveling amyloid toxicity pathway in NIH3T3 cells by a combined proteomic and¹H‐NMR metabonomic approach

Cited by 10 publications

References 72 publications

Glycation Accelerates Fibrillization of the Amyloidogenic W7FW14F Apomyoglobin

Glycation Accelerates Fibrillization of the Amyloidogenic W7FW14F Apomyoglobin

Platelet‐Activating Factor Mediates the Cytotoxicity Induced by W7FW14F Apomyoglobin Amyloid Aggregates in Neuroblastoma Cells

Glycation of Wild‐Type Apomyoglobin Induces Formation of Highly Cytotoxic Oligomeric Species

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Unraveling amyloid toxicity pathway in NIH3T3 cells by a combined proteomic and1H‐NMR metabonomic approach

Cited by 10 publications

References 72 publications

Glycation Accelerates Fibrillization of the Amyloidogenic W7FW14F Apomyoglobin

Glycation Accelerates Fibrillization of the Amyloidogenic W7FW14F Apomyoglobin

Platelet‐Activating Factor Mediates the Cytotoxicity Induced by W7FW14F Apomyoglobin Amyloid Aggregates in Neuroblastoma Cells

Glycation of Wild‐Type Apomyoglobin Induces Formation of Highly Cytotoxic Oligomeric Species

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Unraveling amyloid toxicity pathway in NIH3T3 cells by a combined proteomic and¹H‐NMR metabonomic approach