Effects of N-terminal Acetylation on the Aggregation of Disease-related α-synuclein Variants

Bell, Rosie; Castellana-Cruz, Marta; Nene, Aishwarya; Thrush, Rebecca J.; Xu, Catherine; Kumita, Janet R.; Vendruscolo, Michele

doi:10.1016/j.jmb.2022.167825

Cited by 16 publications

(15 citation statements)

References 56 publications

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“…The N-terminal acetylated wild-type αS was purified as described previously. − ,, Briefly, αS was produced by cotransforming E. coli with the pT7-7 plasmid and an expression pACYCduet plasmid encoding a yeast N-terminal acetyltransferase (NatB), provided by Dr. Dan Mulvihill, University of Kent, Canterbury, UK .…”

Section: Methodsmentioning

confidence: 99%

Secondary Processes Dominate the Quiescent, Spontaneous Aggregation of α-Synuclein at Physiological pH with Sodium Salts

Horne,

Metrick,

Man

et al. 2023

ACS Chem. Neurosci.

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The accurate recapitulation in an in vitro assay of the aggregation process of α-synuclein in Parkinson's disease has been a significant challenge. As α-synuclein does not aggregate spontaneously in most currently used in vitro assays, primary nucleation is triggered by the presence of surfaces such as lipid membranes or interfaces created by shaking, to achieve aggregation on accessible time scales. In addition, secondary nucleation is typically only observed by lowering the pH below 5.8. Here we investigated assay conditions that enables spontaneous primary nucleation and secondary nucleation at pH 7.4. Using 400 mM sodium phosphate, we observed quiescent spontaneous aggregation of α-synuclein and established that this aggregation is dominated by secondary processes. Furthermore, the presence of potassium ions enhanced the reproducibility of quiescent αsynuclein aggregation. This work provides a framework for the study of spontaneous α-synuclein aggregation at physiological pH.

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Section: Methodsmentioning

confidence: 99%

Secondary Processes Dominate the Quiescent, Spontaneous Aggregation of α-Synuclein at Physiological pH with Sodium Salts

Horne,

Metrick,

Man

et al. 2023

ACS Chem. Neurosci.

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“…Various PTMs regulate α-synuclein susceptibility to aggregation by changing its conformation. PTMs such as phosphorylation, ubiquitination, acetylation, nitration, SUMOylation, etc., modify α-synuclein physicochemical properties and affect the aggregation process in synucleinopathies [ 11 , 12 , 29 , 31 , 32 , 33 , 34 ]. α-Synuclein is acetylated constitutively at its N-terminal ( Figure 1 ).…”

Section: α-Synuclein Misfolding Aggregation and Fibrillationmentioning

confidence: 99%

“…α-Synuclein is acetylated constitutively at its N-terminal ( Figure 1 ). As Bell et al recently demonstrated [ 11 , 12 ] this PTM changes the charge and spatial structure of α-synuclein and affects its aggregation and interaction with lipid membranes. Even subtle disturbances caused by PTMs as well as mutations, can lead to remarkable alterations in the aggregation performance of this protein [ 11 , 12 ].…”

Section: α-Synuclein Misfolding Aggregation and Fibrillationmentioning

confidence: 99%

“…Some missense mutations predisposing to diseases (A 53 E, A 53 T, A 30 P, E 46 K, H 50 Q, T 72 M, and E 83 Q) are displayed as blue letters above and under the α-synuclein image (Modified from [ 10 ]). At the upper left corner N-terminal acetylation of α-synuclein is shown, which slows down its aggregation and changes the morphology of the aggregates [ 11 , 12 ].…”

Section: Figurementioning

confidence: 99%

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Synucleins: New Data on Misfolding, Aggregation and Role in Diseases

Surguchov

Surguchev

2022

Biomedicines

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The synucleins are a family of natively unfolded (or intrinsically unstructured) proteins consisting of α-, β-, and γ-synuclein involved in neurodegenerative diseases and cancer. The current number of publications on synucleins has exceeded 16.000. They remain the subject of constant interest for over 35 years. Two reasons explain this unchanging attention: synuclein’s association with several severe human diseases and the lack of understanding of the functional roles under normal physiological conditions. We analyzed recent publications to look at the main trends and developments in synuclein research and discuss possible future directions. Traditional areas of peak research interest which still remain high among last year’s publications are comparative studies of structural features as well as functional research on of three members of the synuclein family. Another popular research topic in the area is a mechanism of α-synuclein accumulation, aggregation, and fibrillation. Exciting fast-growing area of recent research is α-synuclein and epigenetics. We do not present here a broad and comprehensive review of all directions of studies but summarize only the most significant recent findings relevant to these topics and outline potential future directions.

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“…The introduction of lipid vesicles thus not only allows formation of aggregates under quiescent conditions in vitro, but it also provides a simple in vitro model for the interaction of α-synuclein with membranes. The interaction of α-synuclein with lipid-vesicles and their effect on the kinetics of aggregation has been investigated in detail [14,27,28,32,[34][35][36][37][38]. It was thus established, using model membranes, that αsynuclein adsorbs in an α-helical conformation [39] in the head group area and upper acyl layer [40,41] and that protein aggregation is triggered only in situations of protein excess [28,29,42].…”

Section: Introductionmentioning

confidence: 99%

Molecular mechanism of α-synuclein aggregation on lipid membranes revealed

Dear,

Teng,

Ball

et al. 2023

Preprint

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The central hallmark of Parkinson's disease pathology is the aggregation of the α-synuclein protein, which, in its healthy form, is associated with lipid membranes. Purified monomeric α-synuclein is relatively stable in vitro, but its aggregation can be triggered by the presence of lipid vesicles. Despite this central importance of lipids in the context of α-synuclein aggregation, their mechanistic role in this process has not been established to date. Here, we use chemical kinetics to develop a detailed mechanistic model that is able to globally describe the aggregation behaviour of α-synuclein in the presence of DMPS lipid vesicles, across a range of lipid and protein concentrations. Through the application of our kinetic model to experimental data, we find that the reaction is a co-aggregation process involving both protein and lipids and that lipids promote aggregation predominantly by enabling the elongation process. Moreover, we find that the initial formation of aggregates, via primary nucleation, takes place not on the surface of lipid vesicles but at the interfaces present in vitro. Our model will enable mechanistic insights, also in other lipid-protein co-aggregation systems, which will be crucial in the rational design of drugs that inhibit aggregate formation and act at the key points in the α-synuclein aggregation cascade.

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Effects of N-terminal Acetylation on the Aggregation of Disease-related α-synuclein Variants

Cited by 16 publications

References 56 publications

Secondary Processes Dominate the Quiescent, Spontaneous Aggregation of α-Synuclein at Physiological pH with Sodium Salts

Secondary Processes Dominate the Quiescent, Spontaneous Aggregation of α-Synuclein at Physiological pH with Sodium Salts

Synucleins: New Data on Misfolding, Aggregation and Role in Diseases

Molecular mechanism of α-synuclein aggregation on lipid membranes revealed

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