Terai Takuya, Maki Eri, Sugiyama Shigeru, Takahashi Yoshinori, Matsumura Hiroyoshi, Mori Yusuke, Nagano Tetsuo
Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Chem Biol. 2011 Oct 28;18(10):1261-72. doi: 10.1016/j.chembiol.2011.09.007.
Biotin-(strept)avidin complex is widely used in biotechnology because of its extremely high binding constant, but there is no report describing spatiotemporally controlled formation of the complex in live cells. Here, based on X-ray crystal structure analysis and calorimetric data, we designed and synthesized photoreleasable biotins, which show greatly reduced affinity for (strept)avidin, but recover native affinity after UV irradiation. For application at the cell surface, we introduced an amine-reactive moiety into these "caged" biotin molecules. Specific fluorescence imaging of live cells that had been labeled with these agents and then UV-irradiated, was accomplished by addition of streptavidin conjugated with a fluorophore. We also demonstrated the applicability of these compounds for UV-irradiated-cell-specific drug delivery by using caged-biotin-labeled cells, a prodrug, and streptavidin conjugated with a prodrug-activating enzyme.
生物素 -（链霉）抗生物素蛋白复合物因其极高的结合常数而在生物技术中被广泛应用，但尚无关于在活细胞中时空控制该复合物形成的报道。在此，基于X射线晶体结构分析和量热数据，我们设计并合成了光可释放生物素，其对（链霉）抗生物素蛋白的亲和力大大降低，但在紫外线照射后恢复天然亲和力。为了在细胞表面应用，我们将胺反应性部分引入这些“笼化”生物素分子中。通过添加与荧光团偶联的链霉抗生物素蛋白，对用这些试剂标记然后紫外线照射的活细胞进行了特异性荧光成像。我们还通过使用笼化生物素标记的细胞、前药以及与前药激活酶偶联的链霉抗生物素蛋白，证明了这些化合物在紫外线照射细胞特异性药物递送中的适用性。
