Ganglioside GM1 produces stable, short, and cytotoxic Aβ<sub>40</sub>protofibrils

Kumar, Manjeet; Ivanova, Magdalena I.; Ramamoorthy, Ayyalusamy

doi:10.1101/2022.12.28.522135

Cited by 1 publication

(4 citation statements)

References 45 publications

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Section: Stock Preparation Of Aβ Peptides Gm1 and Thtmentioning

confidence: 99%

“…GM1, Aβ42, Aβ40, and ThT stock solutions were prepared and stored as described by Kumar et al, [38]. The critical micellar concentration (CMC) of GM1 was measured using the pyrene fluorescence assay, as reported in our recent study [36,38]. Immediately prior to use, lyophilized peptides and GM1 were dissolved in 20 mM sodium phosphate buffer, pH 7.4, to ~200 µM and 64 µM, respectively.…”

Section: Stock Preparation Of Aβ Peptides Gm1 and Thtmentioning

confidence: 99%

“…To obtain transmission electron microscopic (TEM) images, similar sample preparation, and TEM setting were used as described in our recent study [38]. Briefly, 5 µL sample was transferred to ultrathin holey carbon support film, copper 400 mesh grids, and negatively stained with uranyl acetate (2% w/v; Electron Microscopy Sciences) in water.…”

Section: Transmission Electron Microscopymentioning

confidence: 99%

“…Cell viability assay of Aβ42 amyloid fibrils was performed as described by Kumar et al [38] Briefly, 50 µL differentiation media (Neurobasal-A; Fisher cat# 10888022, B27; Fisher cat# 17-504-044, Fisher GlutaMAX; cat# 35-050-061, Penicillin/Streptomycin) containing 60,000 SH-SY5Y cells/mL (ATCC, catalog # CRL-2266, Lot number: 63724189) were plated in a 96-well plate and differentiated for 10 days with 10 µM retinoic acid (RA) in a humidified incubator at 37 °C with 5% CO2 [41]. Basically, differentiation media was supplemented with RA and was changed every 48 h. On the 11 th day, 50 µL of fresh differentiation media mixed with 50 µL of the samples containing 20 µM Aβ42 amyloid fibrils preformed in the absence and presence of 40 µM GM1, and 40 µM GM1 (alone) was added to the cells and incubated for 48 hours.…”

Section: Cell Viability Assaymentioning

confidence: 99%

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Non-micellar ganglioside GM1 induces an instantaneous conformational change in Aβ₄₂leading to the modulation of the peptide amyloid-fibril pathway

Kumar¹,

Ivanova

Ramamoorthy³

2023

Preprint

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Alzheimer’s disease is a progressive degenerative condition that mainly affects cognition and memory. Recently, distinct clinical and neuropathological phenotypes have been identified in AD. Studies revealed that structural variation in Aβ fibrillar aggregates correlates with distinct disease phenotypes. Moreover, environmental surroundings, including other biomolecules such as proteins and lipids, have been shown to interact and modulate Aβ aggregation. Model membranes containing ganglioside (GM1) clusters are specifically known to promote Aβ fibrillogenesis. This study unravels the modulatory effect of non-micellar GM1, a glycosphingolipid frequently released from the damaged neuronal membranes, on Aβ42amyloid fibril formation. Using far-UV circular dichroism experiments, we observed a spontaneous change in the peptide secondary structure from random-coil to β-turn with subsequent generation of predominantly β-sheet-rich species upon interaction with GM1. Thioflavin-T (ThT) fluorescence assays further indicated that GM1 interacts with the amyloidogenic Aβ42primary nucleus leading to a possible formation of GM1-modified Aβ42fibril. Statistically, no significant difference in toxicity to RA-differentiated SH-SY5Y cells was observed between Aβ42fibrils and GM1-tweaked Aβ42aggregates. Moreover, GM1-modified Aβ42aggregates exhibited prion-like properties in catalyzing the amyloid fibril formation of both major isomers of Aβ, Aβ40, and Aβ42.

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Section: Stock Preparation Of Aβ Peptides Gm1 and Thtmentioning

confidence: 99%