Analysis of Ligand-Receptor Interaction on the Surface of Living Cells by Fluorescence Correlation Spectroscopy
Analysis of Ligand-Receptor Interaction on the Surface of Living Cells by Fluorescence Correlation Spectroscopy
复制标题
通过荧光相关光谱分析活细胞表面的配体-受体相互作用
DOI：
10.2174/1874383801004010028
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
I. Maruyama
中科院分区：
文献类型：
--
作者：
H. Miyagi;I. Maruyama
配体与受体在细胞表面的相互作用传统上是用放射性同位素标记的配体来分析的。为了防止配体-受体复合物的内吞作用，该反应在低温（如4℃）或冰上进行，然后进行大量洗涤以从细胞中去除未结合的配体。为了克服这些缺点，在本研究中，我们采用具有单分子灵敏度的荧光相关光谱（FCS）来分析表皮生长因子（EGF）与其受体EGFR在活细胞表面的相互作用，并成功地确定了它们的解离常数。此外，FCS还被用于测定EGFR在膜中的扩散系数，以及量化细胞表面表达的EGFR分子的数量。
Ligand-receptor interaction on the cell surface has traditionally been analyzed by using radioisotope-labeled ligands. The reaction is carried out at low temperatures such as 4 o C or on ice in order to prevent endocytosis of the ligand- receptor complex, followed by extensive washing to remove unbound ligands from cells. To overcome these drawbacks, in the present study we have employed fluorescence correlation spectroscopy (FCS) with single-molecule sensitivity for the analysis of the interaction of epidermal growth factor (EGF) with its receptor, EGFR, on the surface of living cells, and have successfully determined their dissociation constants. Moreover, FCS has also been used to determine the diffusion coefficient of EGFR in the membrane as well as to quantify the number of EGFR molecules expressed on the cell surface.
影响因子：
3.4
作者：
Vrljic, M;Nishimura, SY;McConnell, HM
通讯作者：
McConnell, HM
DOI：
10.1002/(sici)1097-0320(19990701)36:3
发表时间：
1999-07-01
期刊：
CYTOMETRY
影响因子：
--
作者：
Schwille, P;Korlach, J;Webb, WW
通讯作者：
Webb, WW