Parkhouse R M, Askonas B A
Biochem J. 1969 Nov;115(2):163-9. doi: 10.1042/bj1150163.
Immunoglobulin M biosynthesis was studied with mouse plasma cell tumour MOPC 104E as a model system. Cell suspensions prepared from solid tumours were incubated in vitro with tritiated leucine; the radioactivity incorporated into intracellular and secreted proteins was analysed by polyacrylamide-gel electrophoresis, sucrose-density-gradient centrifugation and precipitation with rabbit antiserum specific for the macroglobulin. The tumour was found to secrete immunoglobulin M and light chains in a 1:2 weight ratio, with lag periods of 20-30min. Within the cells there was a 7s component precipitable with specific antiserum to the macroglobulin that was shown to consist of heavy and light chains. This 7s subunit of the macroglobulin appeared to accumulate in the intracellular environment, so that even after long periods of incubation (3hr.) no more than trace amounts of fully assembled 19s molecules could be detected in cell lysates. Polymerization of the subunits into the pentamer therefore appears to take place shortly before, or simultaneously with, secretion of the molecules.
以小鼠浆细胞瘤MOPC 104E作为模型系统研究了免疫球蛋白M的生物合成。将从实体瘤制备的细胞悬液与氚标记的亮氨酸在体外孵育;通过聚丙烯酰胺凝胶电泳、蔗糖密度梯度离心以及用针对巨球蛋白的兔抗血清沉淀来分析掺入细胞内和分泌蛋白中的放射性。发现该肿瘤以1:2的重量比分泌免疫球蛋白M和轻链,延迟期为20 - 30分钟。在细胞内有一个可被巨球蛋白特异性抗血清沉淀的7s成分,已证明它由重链和轻链组成。巨球蛋白的这个7s亚基似乎在细胞内环境中积累,以至于即使经过长时间孵育(3小时),在细胞裂解物中也只能检测到痕量的完全组装的19s分子。因此,亚基聚合成五聚体似乎在分子分泌前不久或同时发生。