Proteome-Wide Profiling of Cellular Targets Modified by Dopamine Metabolites Using a Bio-Orthogonally Functionalized Catecholamine.
Proteome-Wide Profiling of Cellular Targets Modified by Dopamine Metabolites Using a Bio-Orthogonally Functionalized Catecholamine.
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DOI：
10.1021/acschembio.1c00629
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发表时间：
2021-11-19
影响因子：
4
通讯作者：
Doran, Todd M.
中科院分区：
文献类型：
--
作者：
Hurben, Alexander K.;Erber, Luke N.;Tretyakova, Natalia Y.;Doran, Todd M.
中脑多巴胺能神经元选择性死亡是帕金森病(PD)的一种标志性病理改变，但导致帕金森病神经变性的分子机制尚不清楚。令人信服的证据表明，多巴胺(DA)的失调会诱导神经元应激和损伤反应，这是纹状体退行性变先于帕金森病症状的操作过程。不适当的DA与囊泡的结合增加了胞内DA的水平，这些DA会迅速转化为亲电的多巴奎酮物种(DQs)，这些DQs很容易与蛋白质的亲核体反应，形成共价修饰，从而改变蛋白质的天然结构和功能。这些所谓的DA蛋白加合物(DPA)已被报道在神经毒性中起作用，它们在神经退行性变方面的丰富与帕金森病的临床和病理特征相关联，表明它们在帕金森病的发病机制中起着因果作用。因此，确定DPA的特征是了解中脑多巴胺能神经元在帕金森病中的易感性的关键的第一步。为了帮助实现这一目标，我们在这里报告了一种新型的DA模拟化合物(DAYNE)，它包含一个双正交炔把手，在水溶液中表现出类似于DA的反应性轮廓。通过将与Dayne形成的DPA连接到荧光报告分子，DPA可以在固定细胞和裂解物中显示出来。代恩使受DQ修饰影响的细胞蛋白质及其通过浓缩的生物活性途径的全球图谱成为可能。我们对神经元SH-SY5Y细胞中DPA的蛋白质组学分析表明，对DPA形成敏感的蛋白质存在于整个蛋白质组中，可能影响与PD有关的几个不同的生物学途径，如内质网(ER)应激、细胞骨架不稳定、蛋白毒性和网状蛋白功能。我们证实了一种参与内质网应激途径的蛋白质，蛋白质二硫键异构酶3(PDIA3)，在我们的化学蛋白质组分析中得到了丰富，并被DA功能抑制，这为失调的细胞DA可能诱导或加剧内质网应激提供了证据。因此，Dayne通过能够表征在生理相关的苯二酚暴露下可重复产生的DPA的特征，对PD期间可能观察到的DA毒性提供了新的机械性见解。我们预计，我们设计和应用的这种基于反应性的探针将普遍适用于阐明代谢性苯醌毒性的机制。
Selective death of midbrain dopaminergic neurons is a hallmark pathology of Parkinson’s disease (PD), but the molecular mechanisms that initiate the cascade of events resulting in neurodegeneration in PD remain unclear. Compelling evidence suggests that dysregulation of dopamine (DA) induces neuronal stress and damage responses that are operative processes in striatal degeneration preceding PD-like symptoms. Improper DA sequestration to vesicles raises cytosolic DA levels, which is rapidly converted into electrophilic dopaquinone species (DQs) that react readily with protein nucleophiles forming covalent modifications that alter the native structure and function of proteins. These so-called DA-protein adducts (DPAs) have been reported to play a role in neurotoxicity, and their abundance with respect to neurodegeneration has been linked to clinical and pathological features of PD that suggest that they play a causal role in PD pathogenesis. Therefore, characterizing DPAs is a critical first step in understanding the susceptibility of midbrain dopaminergic neurons during PD. To help achieve this goal, we report here a novel DA-mimetic (DAyne) containing a biorthogonal alkyne handle that exhibits a reactivity profile similar to DA in aqueous buffers. By linking DPAs formed with DAyne to a fluorescent reporter molecule, DPAs were visualized in fixed cells and within lysates. DAyne enabled global mapping of cellular proteins affected by DQ modification and their bioactive pathways through enrichment. Our proteomic profiling of DPAs in neuronal SH-SY5Y cells indicates that proteins susceptible to DPA formation are extant throughout the proteome, potentially influencing several diverse biological pathways involved in PD such as endoplasmic reticulum (ER) stress, cytoskeletal instability, proteotoxicity, and clathrin function. We validated that a protein involved in the ER stress pathway, protein disulfide isomerase 3 (PDIA3), which was enriched in our chemoproteomic analysis, is functionally inhibited by DA, providing evidence that dysregulated cellular DA may induce or exacerbate ER stress. Thus, DAyne provided new mechanistic insights into DA toxicity that may be observed during PD by enabling characterization of DPAs generated reproducibly at physiologically relevant quinone exposures. We anticipate our design and application of this reactivity-based probe will be generally applicable for clarifying mechanisms of metabolic quinone toxicity.
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醌诱导的蛋白质修饰：1,2-苯醌与蛋白质中硫醇基团反应的动力学偏好
影响因子：
7.4
作者：
Li, Yuting;Jongberg, Sisse;Lund, Marianne N.
通讯作者：
Lund, Marianne N.
影响因子：
8.6
作者：
Bruning, John M.;Wang, Yan;England, Pamela M.
通讯作者：
England, Pamela M.
DOI：
10.1126/science.aam9080
发表时间：
2017-09-22
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Burbulla LF;Song P;Mazzulli JR;Zampese E;Wong YC;Jeon S;Santos DP;Blanz J;Obermaier CD;Strojny C;Savas JN;Kiskinis E;Zhuang X;Krüger R;Surmeier DJ;Krainc D
通讯作者：
Krainc D
影响因子：
30.8
作者：
Chang D;Nalls MA;Hallgrímsdóttir IB;Hunkapiller J;van der Brug M;Cai F;International Parkinson's Disease Genomics Consortium;23andMe Research Team;Kerchner GA;Ayalon G;Bingol B;Sheng M;Hinds D;Behrens TW;Singleton AB;Bhangale TR;Graham RR
通讯作者：
Graham RR
DOI：
10.1073/pnas.1402134111
发表时间：
2014-07-08
影响因子：
11.1
作者：
Lohr, Kelly M.;Bernstein, Alison I.;Miller, Gary W.
通讯作者：
Miller, Gary W.