Falach A, Nathan I, Baram S, Porat N, Dvilansky A, Parola A H
Department of Chemistry, Faculty of Natural Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
Bioconjug Chem. 1997 Jul-Aug;8(4):459-65. doi: 10.1021/bc970063f.
A fluorescent analog of human recombinant interferon-gamma (IFN-gamma) was prepared for the first time. The recovered pyrene-labeled IFN-gamma (py-IFN-gamma), with an estimated seven pyrene molecules per IFN-gamma, retained over half of its original biological activity. Binding of py-IFN-gamma to human amnion WISH cells showed appreciable enhancement in fluorescence polarization from 0.055 to 0.215 and in fluorescence lifetime from 56 to 80 ns. The ratio of the vibronic peaks did not change, indicating that the pyrene molecules remained in water environment even after binding. Py-IFN-gamma provides a novel tool for unraveling the mechanism of the initial interaction between this antiproliferative lymphokine and its target, cancer cell membrane receptors. Its fluorescence could provide the means to follow receptor recycling when it occurs.
首次制备了人重组干扰素 -γ（IFN -γ）的荧光类似物。回收的芘标记的 IFN -γ（py - IFN -γ），估计每个 IFN -γ 分子有七个芘分子，保留了其原始生物活性的一半以上。py - IFN -γ 与人羊膜 WISH 细胞的结合显示荧光偏振从 0.055 显著增强至 0.215，荧光寿命从 56 纳秒延长至 80 纳秒。振动峰的比例没有变化，表明芘分子即使在结合后仍处于水环境中。py - IFN -γ 为揭示这种抗增殖淋巴因子与其靶标癌细胞膜受体之间初始相互作用的机制提供了一种新工具。其荧光可以为追踪受体循环发生时的情况提供手段。
