Protein−DNA Double and Triple Layers: Interaction of Biotinylated DNA Fragments with Solid Supported Streptavidin Layers
Protein−DNA Double and Triple Layers: Interaction of Biotinylated DNA Fragments with Solid Supported Streptavidin Layers
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蛋白质-DNA 双层和三层：生物素化 DNA 片段与固体支持的链霉亲和素层的相互作用
DOI：
10.1021/la971352v
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发表时间：
1998
期刊：
影响因子：
3.9
通讯作者：
Michael Strube
中科院分区：
文献类型：
--
作者：
K. Ijiro;H. Ringsdorf;E. Birch‐Hirschfeld;S. Hoffmann;Ute Schilken;Michael Strube
链霉亲和素与生物素化脂类在空气−水界面的特定相互作用导致了光学各向异性的二维链霉亲和素(2-D)晶体的形成，其中原始的四个生物素结合位点中的两个保持自由。这些组装的链霉亲和素基质被用作对接末端或中心位置生物素化的双链寡核苷酸的模板。将生物素化的脂单分子膜沉积在石英晶体微天平(QCM)的电极上，检测到蛋白质与寡核苷酸的对接过程与QCM上质量变化相关的频率变化。双生物素化的双链寡核苷酸结合在链霉亲和素的初级层上，使设计蛋白质−DNA−蛋白质三层成为可能。限制性内切酶的水解表明，与链霉亲和素层结合的生物素标记的DNA仍然具有生物活性。
The specific interaction of streptavidin with biotinylated lipids at the air−water interface leads to a formation of optically anisotropic two-dimensional streptavidin (2-D) crystals, where two of the original four biotin-binding sites remain free. These assembled streptavidin matrixes were used as a template for docking of double-stranded oligonucleotides biotinylated at a terminal or a centered position. A biotinylated lipid monolayer was deposited on an electrode of a quartz crystal microbalance (QCM), and docking processes of the protein and the oligonucleotides were detected as frequency changes related by mass changes on the QCM. The bis-biotinylated double-stranded oligonucleotides bound to the primary streptavidin layers made it possible to engineer protein−DNA−protein triple layers. Hydrolysis by a restriction endonuclease indicates that the biotinylated DNA bound to the streptavidin layers remains bioactive.