Madsen P, Nielsen S, Celis J E
J Cell Biol. 1986 Dec;103(6 Pt 1):2083-9. doi: 10.1083/jcb.103.6.2083.
A monoclonal antibody (mAB 1C4C10) that reacts specifically with human nuclear proteins IEF 8Z30 and 8Z31 (charge variants; HeLa protein catalogue number; Bravo, R., and J. E. Celis, 1982, Clin. Chem., 28:766-781) has been microinjected into the cytoplasm of cultured cells that either express (primates) or lack these proteins (at least having similar molecular weights and pIs; other species), and its cellular localization has been determined by indirect immunofluorescence. Nuclear localization (nucleolar and nucleoplasmic) of the antibody was observed only in cells expressing these antigens, suggesting that a determinant present in IEF 8Z30 and 8Z31 is required for cytoplasm-nuclear translocation. Nuclear migration was not inhibited by cycloheximide, implying that these proteins may shuttle between nucleus and cytoplasm. The results assumed to support the signal rather than the free diffusion model are further supported by microinjection experiments using antibodies (proliferating cell nuclear antigen/cyclin, DNA) that react with nuclear components but do not recognize cytoplasmic antigens. Furthermore, they raise the possibility that some nonnuclear proteins may be transported to the nucleus by interacting with proteins harboring nuclear location signals.
一种单克隆抗体(mAB 1C4C10)能与人核蛋白IEF 8Z30和8Z31(电荷变体;HeLa蛋白目录编号;Bravo,R.,和J.E. Celis,1982,《临床化学》,28:766 - 781)特异性反应。已将该抗体显微注射到表达这些蛋白的培养细胞(灵长类)或缺乏这些蛋白的培养细胞(至少具有相似分子量和等电点;其他物种)的细胞质中,并通过间接免疫荧光确定其细胞定位。仅在表达这些抗原的细胞中观察到抗体的核定位(核仁和核质),这表明IEF 8Z30和8Z31中存在的一个决定簇是细胞质 - 核转运所必需的。环己酰亚胺不抑制核迁移,这意味着这些蛋白可能在细胞核和细胞质之间穿梭。使用与核成分反应但不识别细胞质抗原的抗体(增殖细胞核抗原/细胞周期蛋白、DNA)进行的显微注射实验进一步支持了假设支持信号而非自由扩散模型的结果。此外,这些结果还提出了一种可能性,即一些非核蛋白可能通过与携带核定位信号的蛋白相互作用而被转运到细胞核。