Bhan P, Miller P S
Department of Biochemistry, School of Hygiene and Public Health, Johns Hopkins University, Baltimore, Maryland.
Bioconjug Chem. 1990 Jan-Feb;1(1):82-8. doi: 10.1021/bc00001a011.
The preparation of oligodeoxyribonucleoside methylphosphonates derivatized with 3-[(2-aminoethyl)carbamoyl]psoralen [(ae)CP] is described. These derivatized oligomers are capable of cross-linking with single-stranded DNA via formation of a photoadduct between the furan side of the psoralen ring and a thymidine of the target DNA when the oligomer-target duplex is irradiated with 365-nm light. The photoreactions of (ae)CP-derivatized methylphosphonate oligomers with single-stranded DNA targets in which the position of the psoralen-linking site is varied are characterized and compared to results obtained with oligomers derivatized with 4'-[[N-(aminoethyl)amino]methyl]-4,5',8-trimethylpsoralen [(ae)AMT]. It appears that the psoralen ring can stack on the terminal base pair formed between the oligomer and its target DNA or can intercalate between the last two base pairs of the oligomer-target duplex. Oligomers derivatized with (ae)CP cross-link efficiently to a thymidine located in the last base pair (n position) or 3' to the last base pair (n + 1 position) of the target, whereas the (ae)AMT-derivatized oligomers cross-link most efficiently to a thymidine located in the n + 1 position. The results show that both the extent and kinetics of cross-linking are influenced by the location of the psoralen-linking site in the oligomer-target duplex.
本文描述了用3-[(2-氨基乙基)氨基甲酰基]补骨脂素[(ae)CP]衍生化的寡脱氧核糖核苷甲基膦酸酯的制备方法。当用365nm光照射寡聚物-靶标双链体时，这些衍生化的寡聚物能够通过补骨脂素环的呋喃侧与靶标DNA的胸腺嘧啶形成光加合物，从而与单链DNA交联。对(ae)CP衍生化的甲基膦酸酯寡聚物与单链DNA靶标的光反应进行了表征，其中补骨脂素连接位点的位置是变化的，并与用4'-[[N-(氨基乙基)氨基]甲基]-4,5',8-三甲基补骨脂素[(ae)AMT]衍生化的寡聚物得到的结果进行了比较。似乎补骨脂素环可以堆积在寡聚物与其靶标DNA之间形成的末端碱基对上，或者可以插入寡聚物-靶标双链体的最后两个碱基对之间。用(ae)CP衍生化的寡聚物有效地交联到位于靶标的最后一个碱基对(n位置)或最后一个碱基对的3'端(n + 1位置)的胸腺嘧啶上，而用(ae)AMT衍生化的寡聚物最有效地交联到位于n + 1位置的胸腺嘧啶上。结果表明，交联的程度和动力学都受补骨脂素连接位点在寡聚物-靶标双链体中的位置影响。
