NUCLEIC-ACID BINDING-MOLECULES WITH HIGH-AFFINITY AND BASE SEQUENCE SPECIFICITY - INTERCALATING AGENTS COVALENTLY LINKED TO OLIGODEOXYNUCLEOTIDES
NUCLEIC-ACID BINDING-MOLECULES WITH HIGH-AFFINITY AND BASE SEQUENCE SPECIFICITY - INTERCALATING AGENTS COVALENTLY LINKED TO OLIGODEOXYNUCLEOTIDES
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DOI：
10.1073/pnas.81.11.3297
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发表时间：
1984-01-01
期刊：
影响因子：
--
通讯作者：
HELENE, C
中科院分区：
文献类型：
--
作者：
ASSELINE, U;DELARUE, M;HELENE, C
合成了通过聚甲基连接剂与插层剂共价连接的低聚脱氧核糖核苷酸。通过3'-磷酸基团[(Tp)n(CH2) mAcr]与吖啶染料（Acr）连接的低胸苷酸酯特异性地与互补序列相互作用。这种相互作用被插层剂强烈地稳定下来。利用吸收光谱和荧光光谱分析表明，（Tp）n(CH2) mAcr与poly（rA）之间的络合物形成涉及n个A X T碱基对的形成，其中n为寡核苷酸中胸腺嘧啶的数目。吖啶环插入在A X T碱基对之间。荧光激发光谱揭示了吖啶环存在两种环境，其相对贡献取决于连接体长度(m)。从（Tp）4(CH2) mAcr与聚（rA）的位点结合和沿多核苷酸晶格的寡核苷酸之间的协同相互作用的角度分析了它们的结合。热力学参数表明，共价连接的吖啶环强有力地稳定了寡核苷酸与其互补序列的结合。稳定化取决于连杆的长度；m = 5的化合物比m = 3的化合物更稳定。这些结果为合成对核酸碱基序列具有高亲和力和高特异性的分子家族开辟了道路。
Oligodeoxyribonucleotides covalently linked to an intercalating agent via a polymethylene linker were synthesized. Oligothymidylates attached to an acridine dye (Acr) through the 3'-phosphate group [(Tp)n(CH2) mAcr ] specifically interact with the complementary sequence. The interaction is strongly stabilized by the intercalating agent. By using absorption and fluorescence spectroscopies, it is shown that complex formation between (Tp)n(CH2) mAcr and poly(rA) involves the formation of n A X T base pairs, where n is the number of thymines in the oligonucleotide. The acridine ring intercalates between A X T base pairs. Fluorescence excitation spectra reveal the existence of two environments for the acridine ring, whose relative contributions depend on the linker length (m). The binding of (Tp)4(CH2) mAcr to poly(rA) is analyzed in terms of site binding and cooperative interactions between oligonucleotides along the polynucleotide lattice. Thermodynamic parameters show that the covalent attachment of the acridine ring strongly stabilizes the binding of the oligonucleotide to its complementary sequence. The stabilization depends on the linker length; the compound with m = 5 gives a more stable complex than that with m = 3. These results open the way to the synthesis of a family of molecules exhibiting both high-affinity and high-specificity for a nucleic acid base sequence.