一种改进的荧光共振能量转移方法分析同质二聚体内蛋白质-蛋白质相互作用(英文)  

An Improved Measurement of Fluorescence Resonance Energy Transfer to Analyse Protein-protein Interaction in Protein Homodimer

作　　者：韩福军[1] 罗永峰[1] 徐军[1] 

机构地区：[1]广州医学院广州呼吸疾病研究所

出　　处：《中国生物化学与分子生物学报》2008年第7期619-629,共11页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No30230180)~~

摘　　要：荧光共振能量转移(fluorescence resonance energy transfer ,FRET)技术日益广泛的应用于检测活细胞中分子内和分子间的相互作用.由于FRET仅发生于相互作用的供体和受体,即供体-受体复合物之间,所以检测的FRET信号必须经标准化处理以去除供体受体比例和浓度的影响然后才能够进行FRET的比较研究.由于供体和受体的比例相同,分子内FRET的检测较为简单;而分子间FRET的检测存在更多的不确定因素,导致现有的方法很难精确定量.根据1类特殊的分子间相互作用,同质二聚体的独特特征,推导出供体-受体复合物的含量,进而开发了1种同质二聚体分子间FRET的精确定量的方法,以1种同质二聚体,雌激素受体α(estrogen receptor alpha ,ERα)为供体和受体对,通过和其它的方法比较,证实了该方法用于FRET检测可获得更可靠的结果.Fluorescence resonance energy transfer （FRET） is increasingly used to study inter- and intramolecular interactions in living cells. Since being proportional to the concentration of the donor-acceptor complex, FRET value must be normalized to exclude the influence of the ratio and the concentration of donor and acceptor for comparison. Different from the intra-molecular FRET which is simplified by the fact that the concentration of the donor is equal to that of the acceptor, the inter-molecular FRET is usually too complicated for most existing measurements to quantify exactly. We deduced the exact proportion of the donor-acceptor complex based on a unique characteristic of homodimer, a special kind of the intermolecular interaction, developed an exact quantification measurement of the FRET. We proved the novel method can generate more reliable estimation of FRET value by comparison with other methods using a homodimer, estrogen receptor alpha （ERa）, as a FRET pair.

关 键 词：雌激素受体 荧光共振能量转移 标准化 同质二聚体 蛋白质-蛋白质相互作用 

分 类 号：Q274[生物学—细胞生物学] Q789