Telen M J, Palker T J, Haynes B F
Blood. 1984 Sep;64(3):599-606.
We have previously shown that a murine monoclonal antibody (A3D8) identifies a human erythrocyte protein antigen whose expression is regulated by the Lutheran inhibitor [In(Lu)] gene. In the present study, we demonstrated by immunoprecipitation and Western blot techniques that the antigen defined by A3D8 was on an 80-kD erythrocyte membrane protein. A second 170-kD protein was coprecipitated with the 80-kD protein but failed to show antigen activity by Western blot analysis. The 170-kD protein, when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in two dimensions, was composed of 50- and 30-kD disulfide-linked subunits. In(Lu) Lu[a-b-) erythrocytes differed from Lu(a+b+) or Lu(a-b+) erythrocytes in that In(Lu) deoxycholate erythrocyte membrane extracts contained trace amounts of immunoprecipitable 80-kD protein compared with detergent-solubilized erythrocyte membrane extracts prepared from Lu(a+b+) or Lu(a-b+) subjects.
我们之前已经表明，一种鼠单克隆抗体（A3D8）可识别一种人类红细胞蛋白抗原，其表达受路德抑制因子[In(Lu)]基因调控。在本研究中，我们通过免疫沉淀和蛋白质印迹技术证明，A3D8所定义的抗原存在于一种80-kD的红细胞膜蛋白上。第二种170-kD的蛋白与80-kD的蛋白共沉淀，但通过蛋白质印迹分析未显示出抗原活性。当通过二维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析时，170-kD的蛋白由50-kD和30-kD的二硫键连接亚基组成。In(Lu) Lu(a-b-)红细胞与Lu(a+b+)或Lu(a-b+)红细胞的不同之处在于，与从Lu(a+b+)或Lu(a-b+)受试者制备的去污剂溶解的红细胞膜提取物相比，In(Lu)脱氧胆酸盐红细胞膜提取物中含有痕量的可免疫沉淀的80-kD蛋白。
