Detection and processing of peripheral myelin protein PMP22 in cultured Schwann cells.
Detection and processing of peripheral myelin protein PMP22 in cultured Schwann cells.
复制标题
DOI：
10.1016/s0021-9258(18)82211-6
复制
发表时间：
1993-05
期刊：
影响因子：
--
通讯作者：
S. Pareek;U. Suter;G. Snipes;A. Welcher;E. Shooter;R. A. Murphy
中科院分区：
文献类型：
--
作者：
S. Pareek;U. Suter;G. Snipes;A. Welcher;E. Shooter;R. A. Murphy
外周髓鞘蛋白，22 kDa(PMP22)，是一种与周围神经中的雪旺细胞相关的髓鞘分子(Snipes，G.J.，Suter，U，Welcher，A.和Shooter，E.M.(1992)J.Cell Biol)。117、225-238)。影响PMP22基因的突变与小鼠的颤抖突变有关(Suter，U，Welcher，A.A.，Ozcelik，T.，Snipes，G.J.，Kosaras，B.，Francke，U，Billings-Gagliardi，S.，Sidman，R.L.，and Shooter，E.M.(1992)Nature 356,241-244；苏特，U，莫斯科，J.J.，韦尔彻，A.A.，Snipes，G.J.，Kosaras，B.，Sidman，R.L.，Buchberg，A.M.和Shooter，E.M.(1992)Proc.娜塔莉。阿卡德。SCI。美国A.89,4382-4386)和人类中的夏科-玛丽-牙病(Patel，P.I.，Roa，B.B.，Welcher，A.A.，Schoener-Scott，R.，Traskk，B.J.，Pentao，L.，Snipes，G.J.，Garcia，C.A.，Francke，U，Shooter，E.M.，Lupski，J.R.和Suter，U.(1992)自然基因。1,159-165)。在本报告中，我们研究了培养的大鼠雪旺细胞产生PMP22。雪旺细胞含有编码PMP22的1.8kb的mRNA转录本，其产量在体外受到Forsklin的上调。代谢标记结合针对PMP22合成肽的抗体的免疫沉淀方法表明，雪旺细胞从18 kDa的前体形式产生蛋白质，该前体形式在翻译后被N-连接的糖基化修饰。在雪旺细胞中也检测到与PMP22抗体反应的第二个分子(分子质量，48 kDa)，但通过脉冲追逐标记确定与PMP22在化学上没有关系。大鼠坐骨神经的代谢标记和纯化的大鼠坐骨神经髓鞘的Western印迹分析表明，PMP22的脱糖作用可产生一种大小与雪旺细胞相似的18 kDa蛋白质。这些结果表明，培养的雪旺细胞可以为研究PMP22的产生和功能提供一个很好的模型，并为该蛋白参与遗传性周围神经病变奠定细胞基础。
Peripheral myelin protein, 22 kDa (PMP22), is a myelin molecule associated with Schwann cells in peripheral nerves (Snipes, G. J., Suter, U., Welcher, A. A., and Shooter, E. M. (1992) J. Cell Biol. 117, 225-238). Mutations affecting the PMP22 gene have been implicated in the trembler mutation in mice (Suter, U., Welcher, A. A., Ozcelik, T., Snipes, G. J., Kosaras, B., Francke, U., Billings-Gagliardi, S., Sidman, R. L., and Shooter, E. M. (1992) Nature 356, 241-244; Suter, U., Moskow, J. J., Welcher, A. A., Snipes, G. J., Kosaras, B., Sidman, R. L., Buchberg, A. M., and Shooter, E. M. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 4382-4386) and Charcot-Marie-Tooth Disease in humans (Patel, P. I., Roa, B. B., Welcher, A. A., Schoener-Scott, R., Trask, B. J., Pentao, L., Snipes, G. J., Garcia, C. A., Francke, U., Shooter, E. M., Lupski, J. R., and Suter, U. (1992) Nature genet. 1, 159-165). In this report, we have studied PMP22 production in cultured rat Schwann cells. Schwann cells contain a 1.8-kilobase mRNA transcript coding for PMP22, and its production is up-regulated in vitro by forskolin. Metabolic labeling combined with immunoprecipitation methods using antibodies raised against synthetic peptides of PMP22 reveal that Schwann cells generate the protein from an 18-kDa precursor form which is post-translationally modified by N-linked glycosylation. A second molecule (molecular mass, 48 kDa) that reacted with PMP22 antibodies was also detected in Schwann cells but is not related chemically to PMP22 as determined by pulse-chase labeling. Metabolic labeling of rat sciatic nerve and Western blot analyses of purified rat sciatic nerve myelin reveal that deglycosylation of PMP22 gives rise to an 18-kDa protein similar in size to that in Schwann cells. These results indicate that cultured Schwann cells may provide a good model in which to investigate the production and function of PMP22 and to establish the cellular basis for the protein's involvement in inherited peripheral neuropathies.