Jankowska-Anyszka Marzena, Nogalski Maciej, Darzynkiewicz Edward
Department of Chemistry, Warsaw University, 02093 Warsaw, Pasteura 1, Poland.
Nucleosides Nucleotides Nucleic Acids. 2005;24(5-7):503-6. doi: 10.1081/ncn-200061782.
Cap binding proteins, which recognize the cap structure present at 5' termini of RNA polymerase II transcripts, have been routinely isolated and purified using affinity resins with mononucleotide cap analogs attached. Here we present a new methodology in which dinucleotide cap analog, m7GpppG, has been linked to the EAH-Sepharose. The method is based on derivatization of 2',3'-cis diol of the second nucleotide within the cap structure, with levulinic acid, and subsequent coupling of resulted acetal through its carboxylic group with aminohexyl-agarose.
帽结合蛋白能够识别RNA聚合酶II转录本5'末端的帽结构，常规方法是使用连接了单核苷酸帽类似物的亲和树脂来分离和纯化它们。在此，我们提出一种新方法，即将二核苷酸帽类似物m7GpppG连接到EAH-琼脂糖上。该方法基于对帽结构中第二个核苷酸的2',3'-顺式二醇用乙酰丙酸进行衍生化，然后将所得缩醛通过其羧基与氨基己基琼脂糖偶联。
