Immunological characterization and localization of the Na+/Ca2(+)- exchanger in bovine retina
Immunological characterization and localization of the Na+/Ca2(+)- exchanger in bovine retina
复制标题
牛视网膜Na /Ca2( )-交换器的免疫学特征和定位
作者：
W. Haase;W. Friese;R. Gordon;H. Muller;N. Cook
从牛视网膜杆外段膜中纯化钠/钙交换体，并用于新西兰白色兔的免疫。制备了多克隆抗体，通过Western印迹法评估，发现其特异性结合230 kDa Na+/Ca 2（+）-交换蛋白。该抗体不与高分子量的“边缘蛋白”结合，从而证明该蛋白与Na+/Ca 2（+）-交换器的杆外段不同。我们用多克隆抗体对牛视网膜交换蛋白进行了免疫组织化学定位。荧光显微镜检查发现感光细胞外节的免疫标记强度很大，而其他视网膜细胞层的结合力很小。用电镜免疫金方法，我们发现特异性抗体结合在视杆细胞外节质膜的细胞外侧。视杆-盘膜、视杆内节和视锥光感受器显示无显著标记。因此，我们得出结论，Na+/Ca 2（+）-交换器主要定位在杆外节质膜，最合适的本地化考虑其在脊椎动物光转导过程中的作用。
The sodium/calcium exchanger was purified from bovine retinal rod outer segment membranes and used for the immunization of New Zealand White rabbits. A polyclonal antibody was produced which was found to bind specifically to the 230 kDa Na+/Ca2(+)-exchanger protein as assessed by Western blotting. The antibody did not bind to the high-molecular- weight “rim protein,” thereby demonstrating that this protein is distinct from the rod outer segment of Na+/Ca2(+)-exchanger. We used the polyclonal antibody for immunohistochemically localizing the exchange protein in bovine retina. Fluorescent light microscopy revealed intensive immunolabeling of the photoreceptor outer segments, whereas other retinal cell layers exhibited minimal binding. Using the electron microscopic immunogold method, we found specific antibody binding to the extracellular side of rod outer segment plasma membrane. Rod disk membranes, rod inner segments, and cone photoreceptors displayed no significant labeling. We therefore conclude that the Na+/Ca2(+)-exchanger is localized primarily in the rod outer segment plasma membrane, the most appropriate localization considering its proposed role in the process of vertebrate phototransduction.
使用单克隆抗体在视杆感光盘膜中组织视紫红质和高分子量糖蛋白。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
MacKenzie,D;Molday,RS
通讯作者：
Molday,RS
影响因子：
4.4
作者：
Polans,AS;Burton,MD
通讯作者：
Burton,MD
DOI：
10.1016/s0021-9258(18)83530-x
发表时间：
1989-04
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
N. Cook;L. Molday;D. Reid;U. Kaupp;R. Molday
通讯作者：
N. Cook;L. Molday;D. Reid;U. Kaupp;R. Molday