Yoshimura K, Negishi T, Kaneko A, Sakamoto Y, Kitamura K, Hosokawa T, Hamaguchi K, Nomura M
Dept. of Physiology, Saitama Medical School, Japan.
Neurosci Res. 1996 May;25(1):41-9. doi: 10.1016/0168-0102(96)01026-7.
Two monoclonal antibodies (mAbs), 58A and 46E, were generated against the major protein P0 of bovine peripheral nervous system myelin (PNSM). The reactivities of the mAbs were assessed by enzyme-linked immunosorbent assay (ELISA), Western blot, and immunohistochemistry. Both mAbs, 58A and 46E, reacted to PNSM of bovine, human, rat and rabbit, but not to chicken PNSM or the brains of rat and rabbit. In the Western blot, these mAbs showed specific binding to bovine P0 as well as deglycosylated P0, but not to myelin-associated glycoprotein (MAG) of bovine spinal cord. The analyses of the lysylendopeptidase-digested peptides of bovine P0 revealed that the epitopes for the mAbs 58A and 46E were located on the amino acid residues 68-79 and 210-216, respectively. Since the mAbs 58A and 46E recognize the extracellular domain and the cytoplasmic domain of P0, respectively, they could be useful for studies on P0's role in myelin formation, its adhesive properties, and functions of the N-terminal extracellular and C-terminal cytoplasmic domains of the protein.
针对牛外周神经系统髓磷脂（PNSM）的主要蛋白P0产生了两种单克隆抗体（mAb），即58A和46E。通过酶联免疫吸附测定（ELISA）、蛋白质印迹法和免疫组织化学评估了这些单克隆抗体的反应活性。58A和46E这两种单克隆抗体均与牛、人、大鼠和兔的PNSM发生反应，但不与鸡的PNSM或大鼠和兔的脑发生反应。在蛋白质印迹法中，这些单克隆抗体显示出与牛P0以及去糖基化的P0特异性结合，但不与牛脊髓的髓鞘相关糖蛋白（MAG）结合。对牛P0经赖氨酰内肽酶消化后的肽段分析表明，单克隆抗体58A和46E的表位分别位于氨基酸残基68 - 79和210 - 216上。由于单克隆抗体58A和46E分别识别P0的细胞外结构域和细胞质结构域，它们可用于研究P0在髓鞘形成中的作用、其黏附特性以及该蛋白N端细胞外结构域和C端细胞质结构域的功能。
