SYNTHESIS AND HIGH-STABILITY OF COMPLEMENTARY COMPLEXES OF N-(2-HYDROXYETHYL)PHENAZINIUM DERIVATIVES OF OLIGONUCLEOTIDES
SYNTHESIS AND HIGH-STABILITY OF COMPLEMENTARY COMPLEXES OF N-(2-HYDROXYETHYL)PHENAZINIUM DERIVATIVES OF OLIGONUCLEOTIDES
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DOI：
10.1021/bc00017a010
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发表时间：
1992-09-01
影响因子：
4.7
通讯作者：
ZARYTOVA, VP
中科院分区：
文献类型：
--
作者：
LOKHOV, SG;PODYMINOGIN, MA;ZARYTOVA, VP
提出了两种合成5‘-和/或3’-端磷酸基团含有N-(2-羟乙基)吩嗪(PHN)残基的寡核苷酸的简单方法。通过十二核苷酸d(PApCpCpTpGpTpTpTpTpGpGpGpTpTpTpGpGpC)、七核苷酸d(PCpCpApApApCpA)和后者的PHN衍生物之间形成的络合物，在寡核苷酸链末端引入一种染料可以强烈地稳定其互补的络合物。测定了络合物形成的T(Max)和热力学参数(DELTAH、DELTAS、DELTAG)。根据这些数据，染料与5‘-末端磷酸基团的偶联是最有利的：与3’-PHN衍生物的2.06+/-0.04kcal/mol相比，DELTAG(37-C)增加了3.59+/-0.04kcal/mol。连接染料和寡核苷酸的连接体从二甲亚甲基延伸到七亚甲基，通常会导致寡核苷酸复合体的不稳定。在所研究的所有双链中，由七核苷酸的3‘，5’-二PHN衍生物形成的互补络合物最稳定。相对于未修饰的络合物，自由能增加了4.96+/-0.04千卡/摩尔。修饰后的络合物在37℃时的缔合常数为9.5.10(6)M-1，而未修饰的络合物的类似值仅为3.10(3)M-1。
Two simple methods for the synthesis of oligonucleotides bearing a N-(2-hydroxyethyl)phenazinium (Phn) residue at the 5'- and/or 3'-terminal phosphate groups are proposed. By forming complexes between a dodecanucleotide d(pApApCpCpTpGpTpTpTpGpGpC), a heptanucleotide d(pCpCpApApApCpA), and Phn derivatives of the latter, it is shown that the introduction of a dye at the end of an oligonucleotide chain strongly stabilizes its complementary complexes. The T(max) and the thermodynamic parameters (DELTAH, DELTAS, DELTAG) of complex formation were determined. According to these data, coupling of a dye with the 5'-terminal phosphate group is the most advantageous: DELTAG(37-degrees-C) is increased by 3.59 +/- 0.04 kcal/mol compared to 2.06 +/- 0.04 kcal/mol for 3'-Phn derivatives. The elongation of the linker, which connects the dye to the oligonucleotide, from a dimethylene up to a heptamethylene usually leads to destabilization of the oligonucleotide complex. The complementary complex formed by the 3',5'-di-Phn derivative of the heptanucleotide was found to be the most stable among all duplexes investigated. Relative to the unmodified complex the increase in free energy was 4.96 +/- 0.04 kcal/mol. The association constant of this modified complex at 37-degrees-C is 9.5.10(6) M-1, whereas the analogous value for the unmodified complex is only 3.10(3) M-1.