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Determination of the stoichiometry, structure, and distribution in living cells of protein complexes from analysis of single-molecular-complexes FRET

机译:通过分析单分子复合物FRET确定蛋白质复合物的化学计量,结构和在活细胞中的分布

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摘要

Advances in two-photon microscopy with spectral resolution (TPM-SR) and the development of a simple theory of Forster Resonance Energy Transfer (FRET) for single molecular complexes recently lead to the development of a novel method for the determination of structure and localization in living cells of membrane protein complexes (Raicu et al., Nature Photon., 3, 2009). An appealing feature of this method is its ability to provide such important information while being unaffected by spurious signals originating from stochastic FRET (Singh and Raicu, Biophys. J., 98, 2010). We will present the results obtained from our recent studies of trimeric FRET calibration standards expressed in the cytoplasm of Chinese hamster ovary (CHO) cells, as well as a model G protein-coupled receptor expressed in the membrane of yeast. Emphasis will be placed on the measurement and analysis of single-molecular-complex FRET data for determination of the quaternary structure of some proteins (or the protein complex structure).
机译:具有光谱分辨率的双光子显微镜技术(TPM-SR)的发展以及单分子复合物简单的Forster共振能量转移(FRET)理论的发展,导致了一种新的结构测定和定位方法的开发。膜蛋白复合物的活细胞(Raicu等人,Nature Photon。,3,2009)。该方法的一个吸引人的特征是它能够提供这样的重要信息,而不受随机FRET产生的虚假信号的影响(Singh和Raicu,Biophys。J.,98,2010)。我们将介绍从我们最近对中国仓鼠卵巢(CHO)细胞质中表达的三聚体FRET校准标准品以及在酵母膜中表达的G型蛋白偶联受体的研究中获得的结果。重点将放在单分子复合物FRET数据的测量和分析上,以确定某些蛋白质的四级结构(或蛋白质复合物结构)。

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