Serial amplification of tau filaments using Alzheimer's brain homogenates and C322A or C322S recombinant tau.

IF 3 4区生物学 Q1 Biochemistry, Genetics and Molecular Biology

FEBS Letters Pub Date : 2025-09-30 DOI:10.1002/1873-3468.70141

Alessia Santambrogio, Sofia Lövestam, Michael A Metrick, Thomas Löhr, Peifeng Xu, Nicholas C T Gallagher, Bernardino Ghetti, Byron Caughey, Sjors H W Scheres, Michele Vendruscolo

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Abstract

The assembly of tau into amyloid filaments is a hallmark of Alzheimer's disease (AD) and other tauopathies. Cryo-EM revealed the existence of disease-specific tau folds, which are challenging to replicate in vitro. We studied three full-length recombinant 0N3R tau forms (the wild-type and the C322A and C322S variants) using an RT-QuIC assay with brain homogenate seeding. C322A tau formed filaments resembling AD paired helical filaments (PHFs) but with a more open C-shaped core. C322S tau yielded structurally distinct filaments with an ordered C-terminal region. Both mutants seeded further aggregation, whereas the wild-type showed poor reproducibility and mainly unfolded aggregates. These results highlight the importance of optimised conditions to produce disease-relevant tau filaments and aid the development of targeted therapies. Impact statement We investigated the seeded assembly of 0N3R tau and its two mutational variants C322A and C322S, using Alzheimer's disease brain homogenates in a real-time quaking-induced conversion (RT-QuIC) assay. The C322A variant formed filaments partially resembling the AD PHF structure, showing the importance of optimised conditions to produce disease-relevant tau filaments.

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使用阿尔茨海默病脑匀浆和C322A或C322S重组tau蛋白对tau蛋白丝进行序列扩增。

tau蛋白组装成淀粉样蛋白细丝是阿尔茨海默病（AD）和其他tau病的标志。Cryo-EM揭示了疾病特异性tau折叠的存在，这在体外复制是具有挑战性的。我们使用RT-QuIC方法研究了三种全长重组0N3R tau（野生型、C322A和C322S变体）。C322A tau形成的细丝类似于AD配对的螺旋细丝（phf），但具有更开放的c形核心。C322S tau产生具有有序c端区域的结构明显的细丝。两种突变体都进一步聚集，而野生型表现出较差的再现性，主要是未展开的聚集。这些结果强调了优化条件以产生与疾病相关的tau蛋白细丝的重要性，并有助于开发靶向治疗。我们研究了0N3R tau及其两个突变变体C322A和C322S的种子组装，使用阿尔茨海默病脑匀浆进行实时振动诱导转化（RT-QuIC）试验。C322A变体形成的细丝部分类似于AD PHF结构，这表明优化条件对于产生与疾病相关的tau细丝的重要性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

FEBS Letters 生物-生化与分子生物学

CiteScore

7.00

自引率

2.90%

发文量

303

审稿时长

1.0 months

期刊介绍： FEBS Letters is one of the world''s leading journals in molecular biology and is renowned both for its quality of content and speed of production. Bringing together the most important developments in the molecular biosciences, FEBS Letters provides an international forum for Minireviews, Research Letters and Hypotheses that merit urgent publication.