Cellular Consequences of Copper Complexes Used To Catalyze Bioorthogonal Click Reactions
Cellular Consequences of Copper Complexes Used To Catalyze Bioorthogonal Click Reactions
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DOI：
10.1021/ja2083027
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发表时间：
2011-11-09
影响因子：
15
通讯作者：
Pezacki, John Paul
中科院分区：
文献类型：
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作者：
Kennedy, David C.;McKay, Craig S.;Pezacki, John Paul
铜的毒性是铜(I)催化的叠氮化-炔环加成(CuAAC)铜基催化剂在生命体系中应用的一个关键问题。铜对哺乳动物细胞的影响和相关毒性取决于配基环境。铜的络合物具有很强的毒性，可以引起细胞新陈代谢的变化；迅速被细胞吸收，所有这些都会影响它们在生命系统中作为CuAAC的催化剂的能力。在此，我们通过测量来评估一些典型的铜配合物在四种人类细胞系上催化CuAAC反应的效果。基于线粒体活性的四甲基偶氮唑盐(3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazolium)研究毒性，电感耦合等离子体质谱研究细胞摄取，相干反斯托克斯拉曼散射(CARS)显微镜研究对脂质代谢的影响。我们发现配体：铜周围的环境影响这三个参数。有趣的是，对于铜(II)-双-L-组氨酸络合物(Cu(His)(2))，在微摩尔浓度下，72 h后观察到细胞摄取和代谢的变化，没有毒性。此外，我们还表明，在其他铜配合物杀死人肝癌细胞的条件下，铜(I)-L-组氨酸是一种有效的催化剂，用于在细胞表面表达的糖基化蛋白中代谢掺入炔基标记的糖(Ac(4)Mannal)后标记活细胞。这一结果表明，铜(组氨酸)2或其衍生物具有潜在的体内应用，其中毒性和催化活性是成功的生物偶联反应的关键因素。
Copper toxicity is a critical issue in the development of copper-based catalysts for copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reactions for applications in living systems. The effects and related toxicity of copper on mammalian Cells are dependent on the ligand environment. Copper complexes can be highly toxic, can induce changes in cellular metabolism; and can be. rapidly taken up by cells all of which can affect their ability to function as catalysts for CuAAC in living systems.. Herein, we have evaluated the effects Of a number of copper complexes that are typically Used, to catalyze CuAAC reactions on four human cell lines by measuring. mitochondrial activity based on the Metabolism of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to study toxicity, inductively coupled plasma mass spectrometry to study cellular uptake, and coherent anti Stokes Raman scattering (CARS) microscopy to study effects on lipid metabolism. We find that ligand:environment around copper influences all three parameters. Interestingly,for the Cu(II)-bis-L-histidine complex (Cu(his)(2)), cellular uptake and metabolic changes are observed with no toxicity after 72 h at micromolar concentrations. Furthermore, we show that under conditions where other copper complexes kill human hepatoma cells, Cu(I)-L-histidine is an effective catalyst for CuAAC labeling of live cells following metabolic incorporation of an alkyne-labeled sugar (Ac(4)ManNAl) into glycosylated proteins expressed on the cell surface. This result suggests that Cu(his)2 or derivatives thereof have potential for in vivo applications where toxicity as well as catalytic activity are critical factors for successful bioconjugation reactions.