Ehrick Robin S, Capaccio Marcello, Puleo David A, Bachas Leonidas G
Department of Chemistry, University of Kentucky, Lexington, Kentucky 40506-0055, USA.
Bioconjug Chem. 2008 Jan;19(1):315-21. doi: 10.1021/bc700196q. Epub 2007 Nov 15.
An increase in bone resorption is one of the main symptoms of osteoporosis, a disease that affects more and more individuals every day. Bisphosphonates are known to inhibit bone resorption and thus are being used as a treatment for osteoporosis. Aminobisphosphonates present a functionality that can be easily used for conjugation to other molecules, such as peptides, proteins, and ligands for protein recognition. In this study, an aminobisphosphonate conjugated with biotin was used as a model linker for protein attachment to bone. With this system, the interaction of biotinylated aminobisphosphonate with hydroxyapatite, a major mineral component of bone, was investigated. Quantification of the binding of aminobisphosphonate to hydroxyapatite was performed using a fluorescently labeled antibody for biotin. Additionally, the interaction of the biotinylated aminobisphosphonate with multiple treatments of cortical bone from the midshaft of a cow femur was studied. It was demonstrated that modified aminobisphosphonate reagents can bind hydroxyapatite and bone at high levels, while the biotin functionality is free to be recognized by the fluorescently labeled antibiotin antibody, suggesting that modified aminobisphosphonates could be used to link other peptides or proteins to the bone surface.
骨吸收增加是骨质疏松症的主要症状之一，这种疾病每天影响着越来越多的人。已知双膦酸盐可抑制骨吸收，因此被用作治疗骨质疏松症的药物。氨基双膦酸盐具有一种功能，可轻松用于与其他分子共轭，如肽、蛋白质以及用于蛋白质识别的配体。在本研究中，一种与生物素共轭的氨基双膦酸盐被用作蛋白质附着于骨的模型连接体。利用该系统，研究了生物素化氨基双膦酸盐与骨的主要矿物质成分羟基磷灰石之间的相互作用。使用针对生物素的荧光标记抗体对氨基双膦酸盐与羟基磷灰石的结合进行定量。此外，还研究了生物素化氨基双膦酸盐与来自奶牛股骨中段的皮质骨多次处理后的相互作用。结果表明，修饰后的氨基双膦酸盐试剂可高水平结合羟基磷灰石和骨，而生物素功能可被荧光标记的抗生物素抗体自由识别，这表明修饰后的氨基双膦酸盐可用于将其他肽或蛋白质连接到骨表面。
