Carey J, Cameron V, de Haseth P L, Uhlenbeck O C
Biochemistry. 1983 May 24;22(11):2601-10. doi: 10.1021/bi00280a002.
The interaction between phage R17 coat protein and its RNA binding site for translational repression was studied as an example of a sequence-specific RNA--protein interaction. Nuclease protection and selection experiments define the binding site to about 20 contiguous nucleotides which form a hairpin. A nitrocellulose filter retention assay is used to show that the binding between the coat protein and a synthetic 21-nucleotide RNA fragment conforms to a simple bimolecular reaction. Unit stoichiometry and a Kd of about 1 nM are obtained at 2 degrees C in buffer containing 0.19 M salt. The interaction is highly sequence specific since a variety of RNAs failed to compete with the 21-nucleotide fragment for coat protein binding.
以噬菌体R17外壳蛋白与其用于翻译抑制的RNA结合位点之间的相互作用为例，研究了序列特异性RNA-蛋白质相互作用。核酸酶保护和筛选实验将结合位点确定为大约20个连续的核苷酸，这些核苷酸形成一个发夹结构。使用硝酸纤维素滤膜滞留测定法表明，外壳蛋白与合成的21个核苷酸RNA片段之间的结合符合简单的双分子反应。在含0.19 M盐的缓冲液中于2℃下获得单位化学计量比和约1 nM的解离常数（Kd）。由于多种RNA无法与21个核苷酸片段竞争外壳蛋白结合，因此这种相互作用具有高度的序列特异性。
