PDI抑制α-synuclein纤维化聚集作用机制研究
发布时间：2019-04-24 19:57
【摘要】：天然无结构蛋白?-synuclein在帕金森症(PD)患者脑部的路易小体中异常聚集,被认为是引起PD的重要原因之一,但是目前关于?-synuclein的聚集机制仍没有定论.蛋白质二硫键异构酶(PDI)是细胞内质网中重要的分子伴侣蛋白,能够阻止内质网中无结构蛋白的聚集.在PD患者的脑细胞内发现PDI过量表达,且酶活性位点半胱氨酸被亚硝基化使其活性受到抑制.体外实验证明,PDI能够抑制?-synuclein的聚集,但其具体的分子机制还不清楚,研究PDI抑制?-synuclein聚集的具体机制可能对于PD治疗有重要意义.该文利用核磁共振(NMR)方法研究了?-synuclein与PDI的相互作用,发现?-synuclein与PDI的结合位点位于?-synuclein的N端;将PDI所有的6个半胱氨酸突变成丝氨酸,得到突变体PDI C-S,发现?-synuclein与PDI C-S的结合位点则位于其C末端;荧光实验结果表明突变体PDI C-S对?-synuclein纤维化聚集的抑制作用减弱,说明PDI抑制?-synuclein的纤维化聚集主要是通过与?-synuclein的N端残基结合来实现的.
[Abstract]:The abnormal aggregation of natural structural protein-synuclein in the Louis bodies of the brain of Parkinson's disease patients with (PD) is considered to be one of the important causes of PD. However, the mechanism of the aggregation of?-synuclein is still unknown. Protein disulfide isomerase (PDI) is an important molecular chaperone protein in the endoplasmic reticulum, which can prevent the aggregation of unstructured proteins in the endoplasmic reticulum. Over-expression of PDI was found in the brain cells of PD patients, and cysteine, the enzyme activity site, was inhibited by nitrosation. It has been proved in vitro that PDI can inhibit the aggregation of?-synuclein, but its molecular mechanism is still unclear. The study of the mechanism of PDI inhibiting?-synuclein aggregation may be of great significance for the treatment of PD. The interaction between?-synuclein and PDI was studied by nuclear magnetic resonance (NMR). It was found that the binding site between-synuclein and PDI was at the N-terminal of?-synuclein. All six cysteine mutants of PDI were transformed into serine, and the mutant PDI Cass S was obtained. It was found that the binding site of?-synuclein to PDI Cass was located at the C-terminal of the mutant. The results of fluorescence assay showed that the inhibitory effect of mutant PDI C _ (S) on fibrosis aggregation of?-synuclein was weakened, indicating that the inhibition of PDI on fibrosis aggregation of?-synuclein was mainly achieved by binding to N-terminal residues of?-synuclein.
【作者单位】： 中国科学院生物磁共振分析重点实验室 波谱与原子分子物理国家重点实验室 武汉磁共振中心(中国科学院武汉物理与数学研究所);中国科学院大学;
【基金】：国家自然科学基金资助项目(21203243)
【分类号】：R742.5
本文编号：2464742
[Abstract]:The abnormal aggregation of natural structural protein-synuclein in the Louis bodies of the brain of Parkinson's disease patients with (PD) is considered to be one of the important causes of PD. However, the mechanism of the aggregation of?-synuclein is still unknown. Protein disulfide isomerase (PDI) is an important molecular chaperone protein in the endoplasmic reticulum, which can prevent the aggregation of unstructured proteins in the endoplasmic reticulum. Over-expression of PDI was found in the brain cells of PD patients, and cysteine, the enzyme activity site, was inhibited by nitrosation. It has been proved in vitro that PDI can inhibit the aggregation of?-synuclein, but its molecular mechanism is still unclear. The study of the mechanism of PDI inhibiting?-synuclein aggregation may be of great significance for the treatment of PD. The interaction between?-synuclein and PDI was studied by nuclear magnetic resonance (NMR). It was found that the binding site between-synuclein and PDI was at the N-terminal of?-synuclein. All six cysteine mutants of PDI were transformed into serine, and the mutant PDI Cass S was obtained. It was found that the binding site of?-synuclein to PDI Cass was located at the C-terminal of the mutant. The results of fluorescence assay showed that the inhibitory effect of mutant PDI C _ (S) on fibrosis aggregation of?-synuclein was weakened, indicating that the inhibition of PDI on fibrosis aggregation of?-synuclein was mainly achieved by binding to N-terminal residues of?-synuclein.
【作者单位】： 中国科学院生物磁共振分析重点实验室 波谱与原子分子物理国家重点实验室 武汉磁共振中心(中国科学院武汉物理与数学研究所);中国科学院大学;
【基金】：国家自然科学基金资助项目(21203243)
【分类号】：R742.5
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