Department of Chemistry, University of Rochester, Rochester, New York 14627, USA.
J Am Chem Soc. 2010 Feb 17;132(6):1752-3. doi: 10.1021/ja909106p.
We demonstrate that Zn(II) porphyrin in Zn(II)cytochrome c (Zn cyt c) is a fluorescence resonance energy transfer (FRET) donor to an Alexa660 dye acceptor. The energy transfer efficiency is dependent on the distance between the two fluorophores as shown through protein denaturation studies of five Zn cyt c variants labeled with Alexa660 in different positions. The relative quantum yield, excitation and emission energies, and labeling efficiencies of this donor-acceptor pair allow for a method of analysis based on sensitized emission of the acceptor. These studies show that Zn(II) porphyrin is an effective energy donor for measurement of molecular-scale distances by FRET.
我们证明，锌(II)原卟啉在锌(II)细胞色素 c（Zn cyt c）中是荧光共振能量转移（FRET）的供体，可向 Alexa660 染料受体转移能量。能量转移效率取决于两个荧光团之间的距离，这可以通过对五个在不同位置用 Alexa660 标记的 Zn cyt c 变体进行蛋白质变性研究来证明。该供体-受体对的相对量子产率、激发和发射能量以及标记效率允许基于受体的敏化发射进行分析。这些研究表明，锌(II)卟啉是通过 FRET 测量分子尺度距离的有效能量供体。
