...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Identification of endothelin 1 and big endothelin 1 in secretory vesicles isolated from bovine aortic endothelial cells.
【24h】

Identification of endothelin 1 and big endothelin 1 in secretory vesicles isolated from bovine aortic endothelial cells.

机译：从牛主动脉内皮细胞中分离出分泌小泡中的内皮素1和大内皮素1的鉴定。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Vesicles containing endothelin 1 (ET-1) were isolated from bovine aortic endothelial cells (BAECs) by fractionation of homogenates on sucrose density gradients by ultracentrifugation. The vesicles were localized at the 1.0/1.2 M sucrose interface using a specific anti-ET-1-(16-21) RIA. Identification of ET-1 and big ET-1 in this fraction was confirmed by HPLC analysis combined with RIA. Morphological examination of the ET-1-enriched fraction by electron microscopy identified clusters of vesicles approximately 100 nm in diameter. Immunostaining of ultrathin cryosections prepared from the vesicle fraction for ET-1 or big ET-1 showed clusters of 15-nm gold particles attached to or within vesicles. Immunofluorescence staining of whole BAECs using a specific ET-1-(16-21) IgG purified by affinity chromatography revealed punctate granulation of the cell cytoplasm viewed under light microscopy. This distinct pattern of staining was shown by confocal light microscopy to be intracellular. Immunofluorescence staining of whole cells with a polyclonal antiserum for big ET-1-(22-39) showed a defined perinuclear localization of precursor molecule. Hence, several different approaches have demonstrated that ET-1 and big ET-1 are localized within intracellular vesicles in BAECs, suggesting that these subcellular compartments are an important site for processing of big ET-1 by endothelin-converting enzyme.
机译：通过超速离心从蔗糖密度梯度上的匀浆液分级分离，从牛主动脉内皮细胞（BAEC）中分离出含有内皮素1（ET-1）的囊泡。使用特定的抗ET-1-（16-21）RIA将囊泡定位在1.0 / 1.2 M蔗糖界面上。通过HPLC分析结合RIA确认了该馏分中的ET-1和大ET-1。通过电子显微镜对富含ET-1的馏分进行形态学检查，鉴定出直径约100 nm的囊泡簇。由ET-1或大ET-1的囊泡级分制备的超薄冷冻切片的免疫染色显示，附着在囊泡上或囊泡内的15 nm金粒子簇。使用亲和层析纯化的特异ET-1-（16-21）IgG对整个BAEC进行免疫荧光染色，可见在光学显微镜下观察到的细胞质点状颗粒化。共聚焦光学显微镜显示这种明显的染色模式是细胞内的。用大ET-1-（22-39）的多克隆抗血清对全细胞进行免疫荧光染色，显示前体分子的明确核周定位。因此，几种不同的方法已证明ET-1和大ET-1位于BAECs的细胞内囊泡中，表明这些亚细胞区室是内皮素转化酶处理大ET-1的重要部位。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号