G82S RAGE polymorphism influences amyloid-RAGE interactions relevant in Alzheimer's disease pathology.
G82S RAGE polymorphism influences amyloid-RAGE interactions relevant in Alzheimer's disease pathology.
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DOI：
10.1371/journal.pone.0225487
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发表时间：
2020
期刊：
影响因子：
3.7
通讯作者：
Rani P
中科院分区：
文献类型：
--
作者：
C RC;Lukose B;Rani P
晚期糖基化终产物受体(RAGE)具有结合淀粉样β蛋白(Aβ42)和介导炎性反应的能力，参与阿尔茨海默病(AD)的病理生理过程。G82S RAGE基因多态性与阿尔茨海默病相关，但这种关联的分子机制尚不清楚。我们先前的计算机研究表明，突变的G82S RAGE具有更高的结合亲和力，这可能是由于N81位N连接糖基化的变化所致。为了证实这一假设，本研究利用分子动力学(MD)模拟方法模拟RAGE的野生型(WT)和G82S糖基化结构，以确定RAGE的整体结构变化，并发现其与Aβ42肽的结合效率。MD轨迹的结合口袋分析表明，与WT RAGE相比，突变的G82S糖基化RAGE突变体中的空腔/结合口袋更容易与外部配体接触，这可以增强RAGE对Aβ的亲和力。为了验证上述概念，分别表达重组WT和突变的RAGE突变体的SHSY5Y细胞株与不同浓度的HiLyte Fluor标记的Aβ42进行体外结合研究。采用饱和结合动力学方法测定Aβ42与RAGE结合的Kd值。Aβ42-WT和Aβ42-突变体RAGE结合的Kd值分别为92±40 nM(95%CI-52~152 nM；R2-0.92)和45±20 nM(95%CI-29~64 nM；R2-0.93)。两个突变体的Kd值均为100 nm，表明RAGE是Aβ42的高亲和力受体，突变体RAGE与WT相比具有更高的亲和力。在突变型RAGE表达细胞系中，结合亲和力的改变与炎症通路的激活有关，这为G82S RAGE与阿尔茨海默病的关系提供了一个机制。
Receptor for advanced glycation end products (RAGE) has been implicated in the pathophysiology of Alzheimers disease(AD) due to its ability to bind amyloid-beta (Aβ42) and mediate inflammatory response. G82S RAGE polymorphism is associated with AD but the molecular mechanism for this association is not understood. Our previous in silico study indicated a higher binding affinity for mutated G82S RAGE, which could be caused due to changes in N linked glycosylation at residue N81. To confirm this hypothesis, in the present study molecular dynamics (MD) simulations were used to simulate the wild type (WT) and G82S glycosylated structures of RAGE to identify the global structural changes and to find the binding efficiency with Aβ42 peptide. Binding pocket analysis of the MD trajectory showed that cavity/binding pocket in mutant G82S glycosylated RAGE variants is more exposed and accessible to external ligands compared to WT RAGE, which can enhance the affinity of RAGE for Aβ. To validate the above concept, an in vitro binding study was carried using SHSY5Y cell line expressing recombinant WT and mutated RAGE variant individually to which HiLyte Fluor labeled Aβ42 was incubated at different concentrations. Saturated binding kinetics method was adopted to determine the Kd values for Aβ42 binding to RAGE. The Kd value for Aβ42- WT and Aβ42-mutant RAGE binding were 92±40 nM (95% CI-52 to 152nM; R2-0.92) and 45±20 nM (95% CI -29 to 64nM; R2-0.93), respectively. The Kd value of <100nM observed for both variants implicates RAGE as a high-affinity receptor for Aβ42 and mutant RAGE has higher affinity compared to WT. The alteration in binding affinity is responsible for activation of the inflammatory pathway as implicated by enhanced expression of TNFα and IL6 in mutant RAGE expressing cell line which gives a mechanistic view for the G82S RAGE association with AD.
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影响因子：
3.3
作者：
Daborg, Jonny;von Otter, Malin;Sjolander, Annica;Nilsson, Staffan;Minthon, Lennart;Gustafson, Deborah R.;Skoog, Ingmar;Blennow, Kaj;Zetterberg, Henrik
通讯作者：
Zetterberg, Henrik
影响因子：
2.9
作者：
Krivov, Georgii G.;Shapovalov, Maxim V.;Dunbrack, Roland L., Jr.
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影响因子：
2.2
作者：
Priesnitz, C.;Sperber, S.;Noor, F.
通讯作者：
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影响因子：
2.9
作者：
Miller, Miles C.;Tavares, Rosemarie;Johanson, Conrad E.;Hovanesian, Virginia;Donahue, John E.;Gonzalez, Liliana;Silverberg, Gerald D.;Stopa, Edward G.
通讯作者：
Stopa, Edward G.
影响因子：
82.9
作者：
Deane, R;Yan, SD;Zlokovic, B
通讯作者：
Zlokovic, B