Quantitative detection of the ligand-dependent interaction between the androgen receptor and the co-activator, Tif2, in live cells using two color, two photon fluorescence cross-correlation spectroscopy

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• 作者：Tilman Rosales ; Virginie Georget ; Daniela Malide ; Aleksandr Smirnov ; Jianhua Xu ; Christian Combs ; Jay R. Knutson ; Jean-Claude Nicolas and Catherine A. Royer
• 刊名：European Biophysics Journal
• 出版年：2007
• 出版时间：February, 2007
• 年：2007
• 卷：36
• 期：2
• 页码：153-161
• 全文大小：366 KB

文摘

Two-photon, two-color fluorescence cross-correlation spectroscopy (TPTCFCCS) was used to directly detect ligand-dependent interaction between an eCFP-fusion of the androgen receptor (eCFP-AR) and an eYFP fusion of the nuclear receptor co-activator, Tif2 (eYFP-Tif2) in live cells. As expected, these two proteins were co-localized in the nucleus in the presence of ligand. Analysis of the cross-correlation amplitude revealed that AR was on average 81 % bound to Tif2 in the presence of agonist, whereas the fractional complex formation decreased to 56 % in the presence of antagonist. Residual AR–Tif2 interaction in presence of antagonist is likely mediated by its ligand-independent activation function. These studies demonstrate that using TPTCFCCS it is possible to quantify ligand-dependent interaction of nuclear receptors with co-regulator partners in live cells, making possible a vast array of structure-function studies for these important transcriptional regulators.