A Clickable Oxysterol Photolabel Retains NMDA Receptor Activity and Accumulates in Neurons
A Clickable Oxysterol Photolabel Retains NMDA Receptor Activity and Accumulates in Neurons
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DOI：
10.3389/fnins.2018.00923
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发表时间：
2018-12-06
影响因子：
4.3
通讯作者：
Mennerick, Steven
中科院分区：
文献类型：
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作者：
Chen, Daniel M.;Ziolkowski, Luke;Mennerick, Steven
调节 NMDA 受体功能的氧甾醇类似物是治疗神经精神疾病的候选药物。然而，这些化合物的细胞作用仍不清楚。例如，这些化合物如何在神经元中划分或运输尚不清楚。在这项研究中，我们利用了结合光标记和点击化学的化学生物学方法。我们引入了一种具有生物活性的氧甾醇类似物，其中包含：（1）二氮丙啶基团，允许永久标记细胞靶标，以及（2）炔基团，允许随后使用叠氮共轭荧光团的 Cu2+ 催化环加成进行原位可视化。该类似物在 NMDA 受体上的生理特性与其他氧甾醇类似，包括与其他氧甾醇样化合物的包涵作用。荧光成像显示该类似物通过能量独立机制在神经元的细胞质中广泛积累。总的来说，这项工作引入了一种新的化学生物学方法来研究氧甾醇的作用，并引入了一种可用于氧甾醇进一步细胞生物学和生化研究的工具。
Oxysterol analogs that modulate NMDA receptor function are candidates for therapeutic development to treat neuropsychiatric disorders. However, the cellular actions of these compounds are still unclear. For instance, how these compounds are compartmentalized or trafficked in neurons is unknown. In this study, we utilized a chemical biology approach combining photolabeling and click chemistry. We introduce a biologically active oxysterol analog that contains: (1) a diazirine group, allowing for the permanent labeling of cellular targets, and (2) an alkyne group, allowing for subsequent in situ visualization using Cu2+ catalyzed cycloaddition of an azide-conjugated fluorophore. The physiological properties of this analog at NMDA receptors resemble those of other oxysterols, including occlusion with other oxysterol-like compounds. Fluorescent imaging reveals that the analog accumulates diffusely in the cytoplasm of neurons through an energy-independent mechanism. Overall, this work introduces a novel chemical biology approach to investigate oxysterol actions and introduces a tool useful for further cell biological and biochemical studies of oxysterols.