Hatanaka Y, Hashimoto M, Kanaoka Y
Research Institute for Wakan-Yaku, Toyama Medical and Pharmaceutical University, Japan.
Bioorg Med Chem. 1994 Dec;2(12):1367-73. doi: 10.1016/s0968-0896(00)82088-x.
The synthesis of a p-[(3-trifluoromethyl)diazirine-3-yl]benzoic acid derivative is described as a new carbene generating heterobifunctional cross-linking reagent. The cross-linker carries a biotin moiety in order to make use of avidin-biotin technology for specific manipulation of cross-linked components. To evaluate the ability of this reagent, the inter-subunit cross-linking of egg-white avidin tetramer was investigated. As a typical application of avidin-biotin technology for cross-linking experiments, a chemiluminescent detection method was examined to identify photobiotinylated components. A cross-linked dimeric product with an apparent molecular mass of 38 kDa was clearly visualized by the combined use of a horseradish peroxidase-streptavidin conjugate and a luminol-based chemiluminescent system.
描述了一种对-[(3-三氟甲基)二氮杂环丙烷-3-基]苯甲酸衍生物的合成，该衍生物是一种新型的卡宾生成异双功能交联剂。该交联剂带有生物素部分，以便利用抗生物素蛋白-生物素技术对交联成分进行特异性操作。为了评估该试剂的能力，研究了蛋清抗生物素蛋白四聚体的亚基间交联。作为抗生物素蛋白-生物素技术在交联实验中的典型应用，研究了一种化学发光检测方法以鉴定光生物素化成分。通过联合使用辣根过氧化物酶-链霉抗生物素蛋白缀合物和基于鲁米诺的化学发光系统，清晰地观察到了表观分子量为38 kDa的交联二聚体产物。
