Chaw Y F, Crane L E, Lange P, Shapiro R
Biochemistry. 1980 Nov 25;19(24):5525-31. doi: 10.1021/bi00565a010.
Cross-linked nucleosides have been isolated from formaldehyde-treated ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) by using reverse-phase high-pressure liquid chromatography. Methylene-bridged products containing cytosine, as well as the purines, have been isolated. A combination of ultraviolet and nuclear magnetic resonance (NMR) spectra, pKa values, hydrolysis, and reduction has been used to establish that the cross-links connect the amino groups of the nucleosides involved. For example, the 6-amino functions of two adenosine residues are linked by a methylene bridge to produce a compound which, when dissolved in deuterated dimethyl sulfoxide, displays 11 unsplit resonances in its proton-decoupled 13C NMR spectrum. The procedure reported here is more rapid and less laborious than an earlier one recommended by us for the isolation of cross-linked products from DNA [Dubelman, S., & Shapiro, R. (1977) Nucleic Acids Res. 4, 1815-1827]. This new approach may be of value in the study of other types of cross-linking reactions involving nucleic acids.
通过反相高压液相色谱法，已从甲醛处理过的核糖核酸（RNA）和脱氧核糖核酸（DNA）中分离出交联核苷。已分离出含有胞嘧啶以及嘌呤的亚甲基桥连产物。利用紫外光谱和核磁共振（NMR）光谱、pKa值、水解和还原等方法，确定交联连接了相关核苷的氨基。例如，两个腺苷残基的6-氨基官能团通过亚甲基桥相连，生成一种化合物，该化合物溶解于氘代二甲基亚砜中时，在其质子去耦13C NMR光谱中显示出11个未分裂的共振峰。此处报道的方法比我们之前推荐的从DNA中分离交联产物的方法[Dubelman, S., & Shapiro, R. (1977) Nucleic Acids Res. 4, 1815 - 1827]更快且更省力。这种新方法可能在涉及核酸的其他类型交联反应研究中具有价值。
