Insight into conformational modification of alpha‐synuclein in the presence of neuronal whole cells and of their isolated membranes

Smaldone, Giovanni; Diana, Donatella; Pollegioni, Loredano; Gaetano, Sonia Di; Fattorusso, Roberto; Pedone, Emilia

doi:10.1016/j.febslet.2015.02.012

Cited by 6 publications

(5 citation statements)

References 34 publications

(79 reference statements)

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“…In the cytosol, a-syn is an intrinsically unstructured monomer with a natively unfolded state (Weinreb et al 1996;Kim 1997;Fauvet et al 2012a;Burre et al 2013). a-Syn was later reported to exist as a metastable tetramer (Bartels et al 2011;Wang et al 2011), though other studies reported contrary evidence (Binolfi et al 2012;Fauvet et al 2012b;Burre et al 2013;Smaldone et al 2015) and the formation of multimers is now thought to result from a-syn interacting with lipid membranes (Burre et al 2014) (Fig. 1).…”

Section: A-synucleinmentioning

confidence: 99%

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Antibodies against alpha‐synuclein: tools and therapies

Vaikath

Hmila

Gupta

et al. 2019

Journal of Neurochemistry

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Synucleinopathies including Parkinson's disease, dementia with Lewy bodies and multiple system atrophy are characterized by the abnormal accumulation and propagation of α‐synuclein (α‐syn) pathology in the central and peripheral nervous system as Lewy bodies or glial cytoplasmic inclusions. Several antibodies against α‐syn have been developed since it was first detected as the major component of Lewy bodies and glial cytoplasmic inclusions. Over the years, researchers have generated specific antibodies that alleviate the accumulation of intracellular aggregated α‐syn and associated pathology in cellular and preclinical models of synucleinopathies. So far, antibodies have been the first choice as tools for research and diagnosis and currently, a wide variety of antibody fragments have been developed as an alternative to full‐length antibodies for increasing its therapeutic usefulness. Recently, conformation specific antibody‐based approaches have been found to be promising as therapeutic strategies, both to block α‐syn aggregation and ameliorate the resultant cytotoxicity, and as diagnostic tools. In this review, we summarize different α‐syn specific antibodies and provide their usefulness in tackling synucleinopathies. This article is part of the Special Issue “Synuclein”.

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Section: A-synucleinmentioning

confidence: 99%

“…; Smaldone et al . ) and the formation of multimers is now thought to result from α‐syn interacting with lipid membranes (Burre et al . ) (Fig.…”

Section: α‐Synucleinmentioning

confidence: 99%

Antibodies against alpha‐synuclein: tools and therapies

Vaikath

Hmila

Gupta

et al. 2019

Journal of Neurochemistry

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“…The subcellular localization for aSyn is described to be cytoplasmatic and associated with the outside of the secretory granules ( Acuna et al, 2014 ; Smaldone et al, 2015 ). If aSyn is strictly cytoplasmic, aSyn and IAPP will not be in direct contact with each other.…”

Section: Discussionmentioning

confidence: 99%

Studies on alpha-synuclein and islet amyloid polypeptide interaction

Wang

Bergström

Ingelsson

et al. 2023

Front. Mol. Biosci.

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Introduction: Parkinson’s disease and type 2 diabetes have both elements of local amyloid depositions in their pathogenesis. In Parkinson’s disease, alpha-synuclein (aSyn) forms insoluble Lewy bodies and Lewy neurites in brain neurons, and in type 2 diabetes, islet amyloid polypeptide (IAPP) comprises the amyloid in the islets of Langerhans. In this study, we assessed the interaction between aSyn and IAPP in human pancreatic tissues, both ex vivo and in vitro.Material and Methods: The antibody-based detection techniques, proximity ligation assay (PLA), and immuno-TEM were used for co-localization studies. Bifluorescence complementation (BiFC) was used for interaction studies between IAPP and aSyn in HEK 293 cells. The Thioflavin T assay was used for studies of cross-seeding between IAPP and aSyn. ASyn was downregulated with siRNA, and insulin secretion was monitored using TIRF microscopy.Results: We demonstrate intracellular co-localization of aSyn with IAPP, while aSyn is absent in the extracellular amyloid deposits. ASyn reactivity is present in the secretory granules of β-cells and some α-cells in human islets. The BiFC-expression of aSyn/aSyn and IAPP/IAPP in HEK293 cells resulted in 29.3% and 19.7% fluorescent cells, respectively, while aSyn/IAPP co-expression resulted in ∼10% fluorescent cells. Preformed aSyn fibrils seeded IAPP fibril formation in vitro, but adding preformed IAPP seeds to aSyn did not change aSyn fibrillation. In addition, mixing monomeric aSyn with monomeric IAPP did not affect IAPP fibril formation. Finally, the knockdown of endogenous aSyn did not affect β cell function or viability, nor did overexpression of aSyn affect β cell viability.Discussion: Despite the proximity of aSyn and IAPP in β-cells and the detected capacity of preformed aSyn fibrils to seed IAPP in vitro, it is still an open question if an interaction between the two molecules is of pathogenic significance for type 2 diabetes.

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“…CD spectra in the presence of intact cells were registered using 8 × 10 5 BALBc 3T3 cells or SVT2 cells at different incubation times (0, 10, 30, 60 min and 24 h) in PBS buffer at 20°C. The baseline was corrected by subtracting the spectrum of the cells alone at the same time of incubation [21][22][23].…”

Section: Circular Dichroism Analysesmentioning

confidence: 99%