Itakura Koichi, Furuhata Atsunori, Shibata Noriyuki, Kobayashi Makio, Uchida Koji
Faculty of Education, Aichi University of Education, Kariya, Japan.
Biochem Biophys Res Commun. 2003 Aug 29;308(3):452-7. doi: 10.1016/s0006-291x(03)01412-8.
2-Hydroxyheptanal (2-HH) is one of the reactive aldehyde species generated during the peroxidation of n-6 polyunsaturated fatty acids, such as linoleic and arachidonic acids. Analogous to the Maillard reaction of reducing sugars, 2-HH readily reacts with lysine epsilon-amino groups. In the present study, to define the occurrence of the Maillard reaction-like lysine modification by 2-HH in vivo, we raised a monoclonal antibody directed to a trihydropyridinone (THPO) structure, 1-alkyl-4-butyl-5-pentyl-1,2,6-trihydropyridin-3-one, formed from 2-HH and lysine, and examined the presence of the antigenic structure in the human atherosclerotic aorta. Mice were immunized with the 2-HH-modified keyhole limpet hemocyanin (KLH) as the immunogen. Using a THPO-carrier protein conjugate, we screened the hybridomas and finally obtained a clone that produced the monoclonal antibody 3C8 (mAb3C8). The antibody strongly recognized bovine serum albumin (BSA) treated with 2-HH, but showed no cross-reactivity with BSAs modified with other related aldehydes. By using this antibody, it was revealed that the antigenic structure was indeed present in atherosclerotic lesions of the human aorta.
2-羟基庚醛（2-HH）是在n-6多不饱和脂肪酸（如亚油酸和花生四烯酸）过氧化过程中产生的活性醛类物质之一。与还原糖的美拉德反应类似，2-HH很容易与赖氨酸的ε-氨基发生反应。在本研究中，为了确定体内2-HH引发的类似美拉德反应的赖氨酸修饰的发生情况，我们制备了一种针对由2-HH和赖氨酸形成的三氢吡啶酮（THPO）结构（1-烷基-4-丁基-5-戊基-1,2,6-三氢吡啶-3-酮）的单克隆抗体，并检测了人动脉粥样硬化主动脉中抗原结构的存在情况。用2-HH修饰的钥孔血蓝蛋白（KLH）作为免疫原对小鼠进行免疫。使用THPO-载体蛋白偶联物筛选杂交瘤，最终获得产生单克隆抗体3C8（mAb3C8）的克隆。该抗体强烈识别经2-HH处理的牛血清白蛋白（BSA），但与用其他相关醛修饰的BSA无交叉反应。通过使用该抗体，发现抗原结构确实存在于人的主动脉粥样硬化病变中。
