...
  • 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 >Structure and function in rhodopsin: Asymmetric reconstitution of rhodopsin in liposomes
【24h】

Structure and function in rhodopsin: Asymmetric reconstitution of rhodopsin in liposomes

机译：视紫红质的结构和功能：脂质体中视紫红质的不对称重建

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

We report on preparation of rhodopsin proteoliposomes with the cytoplasmic domain of rhodopsin facing the exterior of the proteoliposomes. Rhodopsin purified from rod outer segments of bovine retinae by immunoaffinity chromatography in octyl glucoside was reconstituted into liposomes prepared from soybean phospholipids by detergent dialysis. The orientation of rhodopsin in the liposomes was determined by susceptibility of its C terminus to papain and the endoproteinase, Asp-N, followed by SDS/PAGE, which showed that the cytoplasmic domain in at least 90% of rhodopsin faced the exterior of the proteoliposomes. By using escape of ~(32)P-KP_i encapsulated in the proteoliposomes as the assay, the half-life of the proteasomes was ≈8 days. After light activation, rhodopsin in proteoliposomes showed the rate of decay of metarhodopsin II and the initial rate of transducin activation comparable with the rates of rhodopsin in rod outer segment membranes. This finding demonstrates the functional capability of rhodopsin in proteoliposomes for kinetic studies of protein-protein interactions.
机译：我们报告视紫红质蛋白脂质体的制备，视紫红质的胞质结构域面向蛋白脂质体的外部。通过免疫亲和层析在辛基葡糖苷中从牛视网膜视杆外部节段纯化的视紫红质，通过去污剂透析，重构为由大豆磷脂制得的脂质体。视紫红质在脂质体中的取向取决于其C末端对木瓜蛋白酶和内蛋白酶Asp-N的敏感性，然后进行SDS / PAGE，这表明视紫红质中至少90％的胞质结构域面对蛋白脂质体的外部。 。通过将包裹在脂质体中的〜（32）P-KP_i逸出作为测定方法，蛋白酶体的半衰期为≈8天。光活化后，蛋白脂质体中的视紫红质显示出视紫红质II的降解速率和转导蛋白的初始活化速率与杆外节膜中的视紫红质速率相当。这一发现证明了视紫红质在脂质体中用于蛋白质-蛋白质相互作用动力学研究的功能。

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