化学
原位
生物物理学
脂质双层
荧光团
细胞生物学
活体细胞成像
磷脂酰肌醇
细胞信号
生物化学
信号
荧光
膜
细胞
信号转导
生物
物理
有机化学
量子力学
作者
Yong-Han Yoon,Park J. Lee,Svetlana Kurilova,Wonhwa Cho
出处
期刊:Nature Chemistry
[Springer Nature]
日期:2011-10-09
卷期号:3 (11): 868-874
被引量:76
摘要
Membrane lipids are dynamic molecules that play important roles in cell signalling and regulation, but an in situ imaging method for quantitatively tracking lipids in living cells is lacking at present. Here, we report a new chemical method of quantitative lipid imaging using sensors engineered by labelling proteins with an environmentally sensitive fluorophore. A prototype sensor for phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P(2))--a key signalling lipid in diverse cellular processes--was generated by covalently attaching a single 2-dimethylamino-6-acyl-naphthalene group to the N-terminal α-helix of the engineered epsin1 ENTH domain, a protein that selectively binds PtdIns(4,5)P(2). The sensor allows robust and sensitive in situ quantitative imaging in mammalian cells, providing new insight into the spatiotemporal dynamics and fluctuation of this key signalling lipid. Application of the sensor to immune cells reveals the presence of a local threshold PtdIns(4,5)P(2) concentration required for triggering phagocytosis. This sensor strategy is generally applicable to in situ quantification of other cellular lipids.
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