Pagès J M, Pagès C, Bernadac A, Prince P
Centre de Biochimie et de Biologie Moléculaire, CNRS, Marseille, France.
Mol Immunol. 1988 Jun;25(6):555-63. doi: 10.1016/0161-5890(88)90077-6.
Nine monoclonal antibodies (MoF 0-8) directed against the native form (trimeric) of outer membrane protein OmpF of Escherichia coli B were obtained and characterized. All these antibodies bind to OmpF porin in intact E. coli B cells but not OmpF from E. coli K-12 cells which only differ at positions 66, 117 and 262 in the sequence. These antibodies exhibit a specificity to the native form, failing to recognize the denatured form in a liquid immunorecognition assay. Four tested antibodies are able to protect against colicin A, a bacteriotoxin using OmpF as receptor. One monoclonal antibody (MoF 0) is specific to the external topology of native porin in the outer membrane and three antibodies could recognize epitopes present in each conformation of subunits of trimer form. It is concluded that the region around the 66th and more probably around the 262nd amino acids are involved in cell-surface exposed epitopes. Moreover, these results support the assumption that the conformation of protruding regions of OmpF from E. coli B and K-12 are different.
获得并鉴定了九种针对大肠杆菌B外膜蛋白OmpF天然形式（三聚体）的单克隆抗体（MoF 0 - 8）。所有这些抗体都能与完整的大肠杆菌B细胞中的OmpF孔蛋白结合，但不能与仅在序列第66、117和262位不同的大肠杆菌K - 12细胞中的OmpF结合。这些抗体对天然形式具有特异性，在液体免疫识别试验中不能识别变性形式。四种测试抗体能够抵御以OmpF为受体的细菌毒素大肠杆菌素A。一种单克隆抗体（MoF 0）对外膜中天然孔蛋白的外部拓扑结构具有特异性，三种抗体能够识别三聚体形式亚基的每种构象中存在的表位。得出的结论是，第66位周围以及更可能是第262位氨基酸周围的区域参与了细胞表面暴露的表位。此外，这些结果支持了大肠杆菌B和K - 12的OmpF突出区域构象不同的假设。
