蛋白质二硫键异构酶与α-突触核蛋白的相互作用及对其聚集的影响
收稿日期: 2022-01-28
网络出版日期: 2022-03-10
基金资助
国家自然科学基金资助项目(21673284);国家自然科学基金资助项目(21773300)
Inhibition of α-Synuclein Aggregation by the Interaction Between Protein Disulfide Isomerase and α-Synuclein
Received date: 2022-01-28
Online published: 2022-03-10
α-突触核蛋白（α-synuclein，αsyn）的错误折叠和聚集是帕金森症的疾病特征.分子伴侣蛋白质二硫键异构酶（PDI）可在体外结合αsyn的N端并抑制其聚集，但PDI的识别机制至今仍不明确.我们通过液体核磁共振（NMR）实验，发现人源PDI b'xa'可结合αsyn的N端区域.此外，硫黄素T（ThT）荧光实验结果表明PDI b'xa'会显著抑制αsyn的聚集.我们进一步利用NMR滴定实验确定了PDI主要通过b'结构域的疏水空腔结合αsyn.最后，我们以此构建了PDI结合αsyn的对接模型，并提出了PDI抑制αsyn聚集的作用机理.这一工作为理解PDI抑制αsyn聚集提供了实验依据.
关键词： 核磁共振(NMR); α-突触核蛋白(αsyn); 蛋白质二硫键异构酶; 相互作用; 纤维化
裴云山 , 张偲 , 刘晓黎 , 成凯 , 张则婷 , 李从刚 . 蛋白质二硫键异构酶与α-突触核蛋白的相互作用及对其聚集的影响[J]. 波谱学杂志, 2022 , 39(4) : 381 -392 . DOI: 10.11938/cjmr20222974
Abnormally misfolded and aggregated α-synuclein (αsyn) is the hallmark of Parkinson's disease (PD). Molecular chaperone protein disulfide isomerase (PDI) has been shown to interact with αsyn and inhibit its aggregation in vitro, but the mechanism for the recognition of αsyn by PDI is not yet clear. Herein, we used nuclear magnetic resonance (NMR) spectroscopy to identify that human PDI b'xa' bound with the N-terminal domain of αsyn, and thioflavin T (ThT) fluorescence assay revealed that b'xa' domain of PDI significantly inhibited αsyn aggregation. Furthermore, by using NMR titration, we observed that PDI bound to αsyn mainly through its hydrophobic cavity of the b' domain. Based on these findings, a docking model of PDI binding with αsyn was established and a possible mechanism of how PDI inhibits αsyn aggregation was proposed. Our work provides experimental evidences for understanding the inhibitory role of PDI in αsyn aggregation.
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