Kellenbach E R, Remerowski M L, Eib D, Boelens R, van der Marel G A, van den Elst H, van Boom J H, Kaptein R
Department of Chemistry, University of Utrecht, The Netherlands.
Nucleic Acids Res. 1992 Feb 25;20(4):653-7. doi: 10.1093/nar/20.4.653.
The synthesis of an oligonucleotide labeled with 13C at the thymine methyl and 15N at the exocyclic amino groups of the cytosines is described. 13CH3I and 15NH4OH were used as sources of the labels. The labeled oligonucleotide was characterized by several NMR techniques. The duplex possesses a labeled functional group in the major groove at every base pair which makes it a very suitable probe for the study of sequence-specific protein-DNA interaction. The labeled thymine methyl group facilitates the detection of hydrophobic contacts with aliphatic side-chains of proteins. This is demonstrated in an NMR study of a complex between the glucocorticoid receptor DNA-binding domain and the labeled oligomer, which revealed a hydrophobic contact between a thymine methyl group and the methyl groups of a valine residue. There are indications for small differences between the solution structure the X-ray structure of the complex.
描述了一种寡核苷酸的合成，该寡核苷酸在胸腺嘧啶甲基处标记有13C，在胞嘧啶的环外氨基处标记有15N。使用13CH3I和15NH4OH作为标记物来源。通过几种核磁共振技术对标记的寡核苷酸进行了表征。该双链体在每个碱基对的大沟中都具有一个标记的官能团，这使其成为研究序列特异性蛋白质-DNA相互作用的非常合适的探针。标记的胸腺嘧啶甲基基团有助于检测与蛋白质脂肪族侧链的疏水接触。这在对糖皮质激素受体DNA结合结构域与标记的寡聚物之间的复合物的核磁共振研究中得到了证明，该研究揭示了胸腺嘧啶甲基基团与缬氨酸残基的甲基之间的疏水接触。有迹象表明该复合物的溶液结构与X射线结构之间存在微小差异。
