Ren J, Goss D J
Department of Chemistry, Hunter College of the City University of New York, NY 10021-5024, USA.
Nucleic Acids Res. 1996 Sep 15;24(18):3629-34. doi: 10.1093/nar/24.18.3629.
In the initiation of protein synthesis, the mRNA 5'-terminal 7-methylguanosine cap structure and several recognition proteins play a pivotal role. For the study of this cap binding reaction, one approach is to use fluorescence spectroscopy. A ribose diol-modified fluorescent cap analog, anthraniloyl-m7GTP (Ant-m7GTP), was designed and synthesized for this purpose. This fluorescent cap analog was found to have a high quantum yield, resistance to photobleaching and avoided overlap of excitation and emission wavelengths with those of proteins. The binding of Ant-m7GTP with wheatgerm initiation factors elF-4F and elF-(iso)4F was determined. The fluorescent cap analog and m7GTP had similar interactions with both cap binding proteins. Fluorescence quenching experiments showed that the microenvironment of Ant-m7GTP when bound to protein was hydrophobic.
在蛋白质合成起始过程中,mRNA 5'-末端的7-甲基鸟苷帽结构和几种识别蛋白起着关键作用。对于这种帽结合反应的研究,一种方法是使用荧光光谱法。为此设计并合成了一种核糖二醇修饰的荧光帽类似物,邻氨基苯甲酰基-m7GTP(Ant-m7GTP)。发现这种荧光帽类似物具有高量子产率、抗光漂白性,并且避免了激发和发射波长与蛋白质的波长重叠。测定了Ant-m7GTP与小麦胚芽起始因子elF-4F和elF-(iso)4F的结合。荧光帽类似物和m7GTP与两种帽结合蛋白具有相似的相互作用。荧光猝灭实验表明,Ant-m7GTP与蛋白质结合时的微环境是疏水的。