Semisynthesis of Fluorescent Metabolite Sensors on Cell Surfaces
Semisynthesis of Fluorescent Metabolite Sensors on Cell Surfaces
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DOI：
10.1021/ja206915m
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发表时间：
2011-10-12
影响因子：
15
通讯作者：
Johnsson, Kai
中科院分区：
文献类型：
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作者：
Brun, Matthias A.;Griss, Rudolf;Johnsson, Kai
由于缺乏合适的传感器来跟踪活细胞中的代谢物、第二信使和神经递质，阻碍了对信号转导和代谢途径的理解。在这里，我们介绍了一类合理设计的半合成荧光传感器蛋白，称为Snifits，用于测量哺乳动物细胞表面的代谢物浓度。功能Snifits通过遗传编码蛋白质支架的两个选择性化学标记反应在活细胞上组装。我们最好的Snifit显示活细胞上的荧光强度比变化显着高于任何以前报道的细胞表面靶向荧光传感器蛋白。这项工作建立了一个普遍适用的和合理的策略，为目标代谢物的细胞表面靶向的荧光传感器蛋白的生成。
Progress in understanding signal transduction and metabolic pathways is hampered by a shortage of suitable sensors for tracking metabolites, second messengers, and neurotransmitters in living cells. Here we introduce a class of rationally designed semisynthetic fluorescent sensor proteins, called Snifits, for measuring metabolite concentrations on the cell surface of mammalian cells. Functional Snifits are assembled on living cells through two selective chemical labeling reactions of a genetically encoded protein scaffold. Our best Snifit displayed fluorescence intensity ratio changes on living cells significantly higher than any previously reported cell-surface-targeted fluorescent sensor protein. This work establishes a generally applicable and rational strategy for the generation of cell-surface-targeted fluorescent sensor proteins for metabolites of interest.