Gaur V P, Adamus G, Arendt A, Eldred W, Possin D E, McDowell J H, Hargrave P A, Sarthy P V
Department of Ophthalmology, University of Washington, Seattle 98195.
Vision Res. 1988;28(7):765-76. doi: 10.1016/0042-6989(88)90023-5.
We have raised monoclonal antibodies to photoreceptor cells in the retina of the turtle (Pseudemys scripta elegans). One of these antibodies, 15-18 (an IgG1), was studied by immunoelectron microscopy using colloidal gold, and found to bind to the outer segments of all rods and some single cones, but did not stain turtle double cones. Immunoblotting and immunoprecipitation show that antibody 15-18 binds to an antigen of apparent Mr approximately 34,5000 which is probably turtle opsin. Antibody 15-18 binds visual pigments from several species, including bovine opsin. In order to determine the antigenic site bound by 15-18 in bovine opsin, synthetic peptides were used as competitors in an enzyme-linked immunoassay (ELISA). The antigenic site is located in the surface loop connecting rhodopsin helices IV-V, in the sequence 190-197. Antibody 15-18 binds to the external surface of rod cell outer segments, thus providing direct evidence for the predicted orientation of rhodopsin in the plasma membrane.
我们制备了针对乌龟（滑龟指名亚种）视网膜光感受器细胞的单克隆抗体。其中一种抗体15 - 18（一种IgG1），通过胶体金免疫电子显微镜进行研究，发现它能与所有视杆细胞的外段以及一些单视锥细胞结合，但不标记乌龟的双视锥细胞。免疫印迹和免疫沉淀表明，抗体15 - 18与一种表观分子量约为34,5000的抗原结合，该抗原可能是乌龟视蛋白。抗体15 - 18能结合多种物种的视觉色素，包括牛视蛋白。为了确定15 - 18在牛视蛋白中结合的抗原位点，在酶联免疫吸附测定（ELISA）中使用合成肽作为竞争剂。抗原位点位于视紫红质螺旋IV - V之间的表面环中，序列为190 - 197。抗体15 - 18与视杆细胞外段的外表面结合，从而为视紫红质在质膜中的预测方向提供了直接证据。
