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Ultrastructural localization of dystrophin in human muscle by using gold immunolabelling.
Proceedings of the Royal Society B: Biological Sciences ( IF 4.7 ) Pub Date : 1990-05-22 , DOI: 10.1098/rspb.1990.0034
M J Cullen 1 , J Walsh , L V Nicholson , J B Harris
Affiliation  

Immunolabelling with a 5 nm gold probe was used to localize dystrophin at the ultrastructural level in human muscle. The primary antibody was monoclonal, raised against a segment (amino acids 1181-1388) from the rod domain of dystrophin. The antibody (Dy4/6D3) is specific for dystrophin and shows no immunoreactivity with any protein from mdx mouse muscle or from patients with a gene deletion spanning part of the molecule recognized by the antibody (Nicholson et al. 1989 a; England et al. 1990). Using this antibody, labelling was almost entirely confined to a narrow 75 nm rim at the periphery of the muscle fibres. Histograms of the distance from the gold probe to the cytoplasmic face of the plasma membrane and of the distance between gold probes (nearest neighbour in a plane parallel with the plasma membrane) displayed modes at approximately 15 nm and 120 nm, respectively. The distribution of the probe was the same in longitudinal and transverse sections of the muscle. These observations suggest that the rod portion of the dystrophin molecule is normally arranged close to the cytoplasmic face of the plasma membrane and that the molecules form an interconnecting network. Labelling was not associated with the transverse tubular system.

中文翻译:

肌营养不良蛋白在人体肌肉中的超微结构定位,采用了金免疫标记。

使用5 nm金探针进行免疫振铃,将肌营养不良蛋白定位于人肌肉的超微结构水平。一抗是单克隆的,针对抗肌萎缩蛋白杆结构域的一个区段(氨基酸1181-1388)产生。抗体(Dy4 / 6D3)对肌营养不良蛋白具有特异性,对mdx小鼠肌肉或基因缺失的患者的任何蛋白质都没有免疫反应,该蛋白质跨越了该抗体识别的分子的一部分(Nicholson等,1989 a; England等。 1990)。使用这种抗体,标记几乎完全局限于肌肉纤维外围的75 nm窄边缘。从金探针到质膜细胞质面的距离的直方图以及金探针之间的距离(与质膜平行的平面中的最近邻居)的直方图分别显示在约15 nm和120 nm处的模式。探针在肌肉的纵向和横向部分的分布是相同的。这些观察结果表明,肌营养不良蛋白分子的杆部分通常布置在质膜的细胞质表面附近,并且该分子形成相互连接的网络。标签与横管系统无关。这些观察结果表明,肌营养不良蛋白分子的杆部分通常布置在质膜的细胞质表面附近,并且该分子形成相互连接的网络。标签与横管系统无关。这些观察结果表明,肌营养不良蛋白分子的杆部分通常布置在质膜的细胞质表面附近,并且该分子形成相互连接的网络。标签与横管系统无关。
更新日期:2019-11-01
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