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利用时间分辨荧光显微镜观察单细胞中的脂质-受体相互作用。

Visualization of lipid-receptor interactions on single cells by time-resolved imaging fluorescence microscopy.

机构信息

Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, P.O. Box 2841, D-37018, Göttingen, Germany.

出版信息

J Fluoresc. 1994 Dec;4(4):295-8. doi: 10.1007/BF01881443.

DOI:10.1007/BF01881443
PMID:24233602
Abstract

The physical interaction between plasma-membrane lipids and the epidermal growth factor (EGF)-receptor was investigated on single A431 human epidermoid carcinoma cells by monitoring fluorescence resonance energy transfer (FRET) between exogeneously added fluorescein-EGF (donor) and 2-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)-1-hexadecanoyl-sn-glycero-3-phosphocholine (Bodipy-PC, acceptor) using donor-photobleaching FRET-microscopy. The measured mean FRET-efficiency of 13% is indicative of such a physical interaction and exemplifies the great potential and sensitivity of time-resolved imaging fluorescence microscopy techniques for the study of lipid-receptor interactions on single cells.

摘要

通过监测荧光共振能量转移(FRET),研究了等离子体膜脂质与表皮生长因子(EGF)受体之间的物理相互作用。在单个 A431 人表皮样癌细胞中,使用外源添加的荧光素-EGF(供体)和 2-(4,4-二氟-5,7-二甲基-4-硼-3a,4a-二氮杂-s-茚并-3-戊酰基)-1-十六酰基-sn-甘油-3-磷酸胆碱(Bodipy-PC,受体)之间的 FRET 显微镜来监测荧光共振能量转移(FRET)。测量的平均 FRET 效率为 13%,表明存在这种物理相互作用,并说明了时间分辨成像荧光显微镜技术在研究单个细胞上的脂质-受体相互作用方面的巨大潜力和敏感性。

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本文引用的文献

1
Resonance energy transfer microscopy: observations of membrane-bound fluorescent probes in model membranes and in living cells.共振能量转移显微镜:模型膜和活细胞中膜结合荧光探针的观察
J Cell Biol. 1986 Oct;103(4):1221-34. doi: 10.1083/jcb.103.4.1221.
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Luminescence digital imaging microscopy.发光数字成像显微镜。
Annu Rev Biophys Biophys Chem. 1989;18:271-308. doi: 10.1146/annurev.bb.18.060189.001415.
3
Fluorescence resonance energy transfer on single living cells. Application to binding of monovalent haptens to cell-bound immunoglobulin E.
单细胞上的荧光共振能量转移。应用于单价半抗原与细胞结合免疫球蛋白E的结合研究。
Biophys J. 1991 Aug;60(2):307-18. doi: 10.1016/S0006-3495(91)82055-0.
4
Time resolved imaging microscopy. Phosphorescence and delayed fluorescence imaging.时间分辨成像显微镜。磷光和延迟荧光成像。
Biophys J. 1991 Dec;60(6):1374-87. doi: 10.1016/S0006-3495(91)82175-0.
5
Rotational mobility of high-affinity epidermal growth factor receptors on the surface of living A431 cells.活的A431细胞表面高亲和力表皮生长因子受体的旋转运动性。
Biochemistry. 1991 Jun 25;30(25):6162-6. doi: 10.1021/bi00239a012.