Fluorescent live-cell imaging of metabolically incorporated unnatural cyclopropene-mannosamine derivatives.
Fluorescent live-cell imaging of metabolically incorporated unnatural cyclopropene-mannosamine derivatives.
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DOI：
10.1002/cbic.201200719
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发表时间：
2013-01-21
期刊：
影响因子：
3.2
通讯作者：
Devaraj, Neal K.
中科院分区：
文献类型：
--
作者：
Cole, Christian M.;Yang, Jun;Seckute, Jolita;Devaraj, Neal K.
有巨大的兴趣在使用生物正交反应成像的非天然建筑模块，代谢纳入生物合成途径。应用包括可视化聚糖，成像标记有非天然氨基酸的蛋白质，监测细胞增殖和跟踪脂质类似物。[1]对于活细胞成像应用，叠氮化物-环辛炔无催化剂的环加成在提供用荧光团标记小叠氮化物标签的快速且生物相容的方法方面是非常有用的。[1a]最近，人们对探索涉及四嗪的生物正交环加成用于活细胞成像应用的兴趣越来越大。[2]四嗪已被证明通过反电子需求狄尔斯-阿尔德反应与各种应变烯烃和炔烃，包括反式-环辛烯，顺丁烯和环辛炔快速反应。这些反应可用于活细胞成像，并且四嗪可以淬灭常用成像探针如BODIPY染料和荧光素的荧光。[3]这导致反应后的荧光响应，其可以改善信号背景比，这对于细胞内活细胞成像应用特别有用。[1b，4]尽管四嗪环加成对于广泛的代谢成像应用来说将是令人兴奋的发展，但四嗪和环烯偶联伙伴的大尺寸限制了它们被掺入小生物活性分子的能力。为了应对这一挑战，我们最近开发了小而稳定的甲基环丙烯作为荧光四嗪的偶联伙伴。[5]这些标签的分子量与叠氮化物的分子量相媲美，并用于对活哺乳动物细胞中的脂质进行荧光成像。然而，我们感兴趣的是甲基环丙烯是否可以取代叠氮化物在代谢成像应用中具有严格的空间限制。在这里，我们证明了非天然的环丙烯甘露糖胺衍生物可用于成像聚糖活的人类癌细胞系。
There is tremendous interest in the use of bioorthogonal reactions for imaging of unnatural building blocks that are metabolically incorporated into biosynthetic pathways. Applications include visualizing glycans, imaging proteins tagged with unnatural amino acids, monitoring cellular proliferation, and tracking lipid analogues.[1] For live-cell imaging applications, the azide–cyclooctyne catalyst-free cycloaddition has been extremely useful in providing a rapid and biocompatible method to label small azide tags with fluorophore.[1a] Recently, there has been growing interest in exploring bioorthogonal cycloadditions involving tetrazines for live-cell imaging applications.[2] Tetrazines have been shown to react rapidly through inverse-electron-demand Diels–Alder reactions with a variety of strained alkenes and alkynes including trans-cyclooctenes, norbornenes, and cyclooctynes. These reactions can be used for live-cell imaging, and tetrazines can quench the fluorescence of commonly used imaging probes such as BODIPY dyes and fluoresceins.[3] This leads to a fluorogenic response after reaction, which can improve signal-to-background, which is particularly useful for intracellular live-cell imaging applications.[1b, 4] Though tetrazine cycloadditions would be exciting developments for a wide array of metabolic imaging applications, the large size of both the tetrazine and cycloalkene coupling partners has limited their ability to be incorporated into small bio-active molecules. In response to this challenge, we recently developed small and stable methylcyclopropenes as coupling partners for fluorogenic tetrazines.[5] The molecular weight of these tags rivaled those of azides and were used to fluorogenically image lipids in live mammalian cells. However, we were interested in whether methylcyclopropenes could substitute for azides in metabolic imaging applications with stringent steric constraints. Here we demonstrate that unnatural cyclopropenemannosamine derivatives can be used to image glycans on live human cancer cell lines.
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