通过表面增强拉曼光谱研究 Hoechst 33258 聚集和与 DNA 的结合,Journal of Raman Spectroscopy

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通过表面增强拉曼光谱研究 Hoechst 33258 聚集和与 DNA 的结合
Journal of Raman Spectroscopy ( IF 2.4 ) Pub Date : 2022-02-06 , DOI: 10.1002/jrs.6319
Adriana Kenđel ₁ , Virna Zavidić ₁ , Snežana Miljanić ₁

Affiliation

使用表面增强拉曼光谱 (SERS) 研究了 Hoechst 33258 (H33258) 与两种合成 DNA 多核苷酸 poly dA-poly dT 和 poly dG-poly dC 以及天然小牛胸腺 (ct) DNA 的相互作用。鉴于各种化学计量依赖性相互作用，在银胶体悬浮液中以 1/1、1/5 和 1/10 的 [H33258]/[DNA] 摩尔比制备复合物。在分析 DNA 多核苷酸的结合模式之前，研究了染料在银胶体中的聚集趋势，浓度范围为 1 × 10 ^-6 –1 × 10 ^-4 摩尔/升。获得了聚合和单体 H33258 物种特有的 SERS 光谱，另外表明了与 DNA 结合的分子的形式。获得的具有同聚多核苷酸的 H33258 的 SERS 光谱表明分子深度插入 AT 小沟，以及通过 GC 大沟进入的 GC 碱基对之间的部分插入。染料的光谱特征在与 ct-DNA 复合物的光谱中占主导地位，很可能是由于长天然核酸的随机 AT 和 GC 序列影响染料的均匀结合，从而影响 H33258/DNA 复合物吸附到增强金属表面。

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Hoechst 33258 aggregation and binding to DNA studied by surface-enhanced Raman spectroscopy

Interactions of Hoechst 33258 (H33258) with two synthetic DNA polynucleotides, poly dA–poly dT and poly dG–poly dC, as well as natural calf thymus (ct) DNA were studied using surface-enhanced Raman spectroscopy (SERS). Given variety of stoichiometry dependent interactions, complexes were prepared in [H33258]/[DNA] molar ratios of 1/1, 1/5, and 1/10 in the silver colloidal suspension. Prior to analysis of binding modes with DNA polynucleotides, aggregation tendency of the dye in the silver colloid was studied in the concentration range 1 × 10⁻⁶–1 × 10⁻⁴ mol/L. The SERS spectra distinctive of aggregated and monomeric H33258 species were obtained, additionally indicating the form of the molecules bound to DNA. The obtained SERS spectra of H33258 with homo-polymeric polynucleotides implied a deep insertion of the molecules into the AT minor groove, and a partial intercalation between the GC base pairs achieved by the access through the GC major groove. Spectral characteristics of the dye dominated in the spectra of the complexes with ct-DNA, most likely due to random AT and GC sequences of the long natural nucleic acid, affecting uniform binding of the dye and consequently the adsorption of the H33258/DNA complex onto the enhancing metallic surface.

更新日期：2022-02-06

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