通过定量荧光共振能量转移分析来测定生化参数的新趋势。
A new trend to determine biochemical parameters by quantitative FRET assays.
作者信息
Liao Jia-yu, Song Yang, Liu Yan
机构信息
Department of Bioengineering, University of California at Riverside, 900 University Avenue, Riverside, CA 92521, USA.
Center for Bioengineering Research, Bourns College of Engineering, University of California at Riverside, 900 University Avenue, Riverside, CA 92521, USA.
出版信息
Acta Pharmacol Sin. 2015 Dec;36(12):1408-15. doi: 10.1038/aps.2015.82. Epub 2015 Nov 16.
Förster resonance energy transfer (FRET) has been widely used in biological and biomedical research because it can determine molecule or particle interactions within a range of 1-10 nm. The sensitivity and efficiency of FRET strongly depend on the distance between the FRET donor and acceptor. Historically, FRET assays have been used to quantitatively deduce molecular distances. However, another major potential application of the FRET assay has not been fully exploited, that is, the use of FRET signals to quantitatively describe molecular interactive events. In this review, we discuss the use of quantitative FRET assays for the determination of biochemical parameters, such as the protein interaction dissociation constant (K(d)), enzymatic velocity (k(cat)) and K(m). We also describe fluorescent microscopy-based quantitative FRET assays for protein interaction affinity determination in cells as well as fluorimeter-based quantitative FRET assays for protein interaction and enzymatic parameter determination in solution.
福斯特共振能量转移（FRET）已在生物和生物医学研究中广泛应用，因为它能够确定1-10纳米范围内的分子或粒子相互作用。FRET的灵敏度和效率强烈依赖于FRET供体与受体之间的距离。从历史上看，FRET分析已被用于定量推导分子距离。然而，FRET分析的另一个主要潜在应用尚未得到充分利用，即利用FRET信号定量描述分子相互作用事件。在本综述中，我们讨论了使用定量FRET分析来测定生化参数，如蛋白质相互作用解离常数（K(d)）、酶促速度（k(cat)）和K(m)。我们还描述了基于荧光显微镜的定量FRET分析，用于测定细胞中的蛋白质相互作用亲和力，以及基于荧光计的定量FRET分析，用于测定溶液中的蛋白质相互作用和酶参数。
Zotero 插件