PDI抑制α-synuclein纖維化聚集作用機制研究
發(fā)布時間：2019-04-24 19:57
【摘要】：天然無結構蛋白?-synuclein在帕金森癥(PD)患者腦部的路易小體中異常聚集,被認為是引起PD的重要原因之一,但是目前關于?-synuclein的聚集機制仍沒有定論.蛋白質二硫鍵異構酶(PDI)是細胞內質網中重要的分子伴侶蛋白,能夠阻止內質網中無結構蛋白的聚集.在PD患者的腦細胞內發(fā)現(xiàn)PDI過量表達,且酶活性位點半胱氨酸被亞硝基化使其活性受到抑制.體外實驗證明,PDI能夠抑制?-synuclein的聚集,但其具體的分子機制還不清楚,研究PDI抑制?-synuclein聚集的具體機制可能對于PD治療有重要意義.該文利用核磁共振(NMR)方法研究了?-synuclein與PDI的相互作用,發(fā)現(xiàn)?-synuclein與PDI的結合位點位于?-synuclein的N端;將PDI所有的6個半胱氨酸突變成絲氨酸,得到突變體PDI C-S,發(fā)現(xiàn)?-synuclein與PDI C-S的結合位點則位于其C末端;熒光實驗結果表明突變體PDI C-S對?-synuclein纖維化聚集的抑制作用減弱,說明PDI抑制?-synuclein的纖維化聚集主要是通過與?-synuclein的N端殘基結合來實現(xiàn)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.
【作者單位】： 中國科學院生物磁共振分析重點實驗室 波譜與原子分子物理國家重點實驗室 武漢磁共振中心(中國科學院武漢物理與數(shù)學研究所);中國科學院大學;
【基金】：國家自然科學基金資助項目(21203243)
【分類號】：R742.5
本文編號：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.
【作者單位】： 中國科學院生物磁共振分析重點實驗室 波譜與原子分子物理國家重點實驗室 武漢磁共振中心(中國科學院武漢物理與數(shù)學研究所);中國科學院大學;
【基金】：國家自然科學基金資助項目(21203243)
【分類號】：R742.5
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1 王紹彬;蛋白質二硫鍵異構酶（PDI）參與調節(jié)朊病毒相關內質網應激及線粒體功能異常的機制研究[D];中國疾病預防控制中心;2012年
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