Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan 430072, China.
National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
J Mol Biol. 2020 Mar 27;432(7):2141-2163. doi: 10.1016/j.jmb.2020.02.013. Epub 2020 Feb 19.
Cells have evolved molecular chaperones that modulate phase separation and misfolding of amyloidogenic proteins to prevent neurodegenerative diseases. Protein disulfide isomerase (PDI), mainly located at the endoplasmic reticulum and also present in the cytosol, acts as both an enzyme and a molecular chaperone. PDI is observed to be S-nitrosylated in the brain of Alzheimer's disease patients, but the mechanism has remained elusive. We herein report that both wild-type PDI and its quadruple cysteine mutant only having chaperone activity, significantly inhibit pathological phosphorylation and abnormal aggregation of Tau in cells, and significantly decrease the mitochondrial damage and Tau cytotoxicity resulting from Tau aberrant aggregation, highlighting the chaperone property of PDI. More importantly, we show that wild-type PDI is selectively recruited by liquid droplets of Tau, which significantly inhibits phase separation and stress granule formation of Tau, whereas S-nitrosylation of PDI abrogates the recruitment and inhibition. These findings demonstrate how phase separation of Tau is physiologically regulated by PDI and how S-nitrosylation of PDI, a perturbation in this regulation, leads to disease.
细胞已经进化出分子伴侣,它们可以调节淀粉样蛋白的相分离和错误折叠,以预防神经退行性疾病。蛋白二硫键异构酶(PDI)主要位于内质网,也存在于细胞质中,它既是一种酶,也是一种分子伴侣。在阿尔茨海默病患者的大脑中观察到 PDI 发生 S-亚硝基化,但机制仍不清楚。我们在此报告,野生型 PDI 及其仅具有伴侣活性的四半胱氨酸突变体,可显著抑制细胞中 Tau 的病理性磷酸化和异常聚集,并显著降低 Tau 异常聚集导致的线粒体损伤和 Tau 细胞毒性,突出了 PDI 的伴侣性质。更重要的是,我们表明野生型 PDI 被 Tau 的液滴选择性募集,这显著抑制 Tau 的相分离和应激颗粒形成,而 PDI 的 S-亚硝基化则消除了募集和抑制。这些发现表明 Tau 的相分离如何被 PDI 生理调节,以及 PDI 的 S-亚硝基化如何导致这种调节失调,从而导致疾病。