The supramolecular structure of the GPCR rhodopsin in solution and native disc membranes
The supramolecular structure of the GPCR rhodopsin in solution and native disc membranes
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DOI：
10.1080/09687860400020291
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发表时间：
2004-11-01
影响因子：
--
通讯作者：
Fotiadis, D
中科院分区：
文献类型：
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作者：
Suda, K;Filipek, S;Fotiadis, D
视紫红质是一种典型的G蛋白偶联受体，密集分布在视杆细胞外节的盘膜上，被认为是一种单体。然而，越来越多的证据表明，许多G-蛋白偶联受体的二聚化和寡聚化，和原子力显微镜图像显示行的视紫红质二聚体在小鼠椎间盘膜。在这项工作中，我们证明了负染色样品的电子显微镜，蓝色的本地和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，化学交联，并通过蛋白水解，本地牛视紫红质主要存在的二聚体和更高的低聚物。这些结果证实了最近的研究结果从原子力显微镜和分子建模的超分子结构和小鼠视紫红质二聚体的包装安排。
Rhodopsin, the prototypical G-protein-coupled receptor, which is densely packed in the disc membranes of rod outer segments, was proposed to function as a monomer. However, a growing body of evidence indicates dimerization and oligomerization of numerous G-protein-coupled receptors, and atomic force microscopy images revealed rows of rhodopsin dimers in murine disc membranes. In this work we demonstrate by electron microscopy of negatively stained samples, blue native- and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, chemical crosslinking, and by proteolysis that native bovine rhodopsin exists mainly as dimers and higher oligomers. These results corroborate the recent findings from atomic force microscopy and molecular modeling on the supramolecular structure and packing arrangement of murine rhodopsin dimers.