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牛外周神经中帕金蛋白的特性分析。

Characterization of parkin in bovine peripheral nerve.

作者信息

Hase Asako, Yamada Hiroki, Arai Ken, Sunada Yoshihide, Shimizu Teruo, Matsumura Kiichiro

机构信息

Department of Neurology and Neuroscience, Teikyo University School of Medicine, 2-11-1 Kaga, Itabashi-ku, Tokyo 173-8605, Japan.

出版信息

Brain Res. 2002 Mar 15;930(1-2):143-9. doi: 10.1016/s0006-8993(02)02241-2.

Abstract

The autosomal recessive juvenile parkinsonism is caused by the mutations of the gene encoding a novel protein called parkin. It has been reported that parkin is expressed in the central nervous system and functions as a ubiquitin-protein ligase (E3) which suppresses neuronal cell degeneration by ubiquitinating misfolded proteins. Thus far, however, it remains unknown if parkin is expressed and functions in the peripheral nervous system. In order to begin to address to this question, we investigated the expression of parkin in bovine peripheral nerve. Reverse transcription polymerase chain reaction analysis demonstrated the presence of parkin transcript in bovine peripheral nerve. The obtained bovine parkin cDNA sequence was identical to that of human except a single nucleotide. Immunoblot analysis demonstrated the expression of parkin protein in bovine peripheral nerve. Immunohistochemical analysis demonstrated the localization of parkin in the axoplasm of myelinated nerve fibers, the Schwann cell cytoplasm and the Schwann cell outer membrane. Furthermore, fractionation analysis indicated the presence of two fractions of parkin in bovine peripheral nerve, the cytosolic fraction and the cell membrane-bound fraction. All together, these results point to diverse roles of parkin in not only the central but also the peripheral nervous system.

摘要

常染色体隐性少年型帕金森病是由一种名为帕金蛋白(parkin)的新型蛋白质编码基因突变引起的。据报道,帕金蛋白在中枢神经系统中表达,并作为一种泛素蛋白连接酶(E3)发挥作用,通过将错误折叠的蛋白质泛素化来抑制神经元细胞变性。然而,到目前为止,帕金蛋白是否在外周神经系统中表达并发挥作用仍不清楚。为了开始解决这个问题,我们研究了帕金蛋白在牛外周神经中的表达。逆转录聚合酶链反应分析表明牛外周神经中存在帕金蛋白转录本。所获得的牛帕金蛋白cDNA序列除了一个单核苷酸外与人类的相同。免疫印迹分析表明牛外周神经中存在帕金蛋白。免疫组织化学分析表明帕金蛋白定位于有髓神经纤维的轴浆、雪旺细胞胞质和雪旺细胞外膜。此外,分级分离分析表明牛外周神经中存在两种帕金蛋白组分,即胞质组分和细胞膜结合组分。总之,这些结果表明帕金蛋白不仅在中枢神经系统而且在外周神经系统中都具有多种作用。

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