Department of Biological Sciences, Sungkyunkwan University, Suwon, 16419, Republic of Korea.
College of Pharmacy, Duksung Women's University, Seoul, 01369, Republic of Korea.
Biochem Biophys Res Commun. 2024 Nov 12;733:150607. doi: 10.1016/j.bbrc.2024.150607. Epub 2024 Aug 24.
3,4-methylenedioxymethamphetamine (MDMA) or publicly known as "ecstasy" is a drug abuse substance. Since antibodies that detect MDMA typically also recognize its chemical analogue, methamphetamine (METH), we identified antibodies specifically recognizing MDMA, but not METH, named 1bB11 and 1bF12, using phage display. The crystal structure of 1bB11 in complex with MDMA was determined at 3.2 Å resolution. Key interactions were found between the epoxide moiety of MDMA and S34 and Y36 of the light chain. Additional interaction with E33 of the heavy chain contributes to anchoring MDMA. Mutagenesis-based biochemical analysis confirmed the importance of these residues in MDMA binding. Comparing the structure of 1bB11 to a scFv6H4, which binds both METH and MDMA, revealed opposite binding orientations. Taken together, our data provides a structural framework for selective binding to MDMA by the 1bB11 antibody, paving a way to develop a highly specific antibody for diagnosis.
3,4-亚甲二氧基甲基苯丙胺（MDMA），俗称“摇头丸”，是一种滥用药物。由于检测 MDMA 的抗体通常也能识别其化学类似物甲基苯丙胺（METH），我们使用噬菌体展示技术鉴定出了特异性识别 MDMA、不识别 METH 的抗体，分别命名为 1bB11 和 1bF12。我们测定了 1bB11 与 MDMA 复合物的晶体结构，分辨率为 3.2 Å。研究发现，MDMA 的环氧化物部分与轻链的 S34 和 Y36 之间存在关键相互作用。重链的 E33 也与 MDMA 发生额外相互作用，有助于固定 MDMA。基于突变的生化分析证实了这些残基在 MDMA 结合中的重要性。将 1bB11 的结构与同时结合 METH 和 MDMA 的 scFv6H4 进行比较，揭示了相反的结合取向。综上所述，我们的数据为 1bB11 抗体对 MDMA 的选择性结合提供了结构框架，为开发用于诊断的高特异性抗体铺平了道路。
