Polans A S, Burton M D, Haley T L, Crabb J W, Palczewski K
R.S. Dow Neurological Sciences Institute, GSH&MC, Portland, OR 97209.
Invest Ophthalmol Vis Sci. 1993 Jan;34(1):81-90.
We investigated the hypothesis that visinin, a cone-specific protein first characterized in chicken retina, is a cone homologue of recoverin and may be the cancer-associated retinopathy (CAR) autoantigen in human cone cells.
Visinin was purified from chicken retinas and tested for binding by CAR antisera. In addition, antibodies specific to visinin were used immunocytochemically and for Western analysis to determine whether visinin is present in human or bovine retinas. Anti-peptide antibodies against recoverin were used immunocytochemically to localize recoverin to mammalian cone cells.
CAR antisera recognized recoverin but not visinin. Furthermore, visinin could not be detected in mammalian retinas by immunocytochemical methods or by attempts to purify the protein. In contrast to visinin, antibodies specific for different regions of the recoverin molecule stained both rod and cone cells in the human retina.
Visinin is not the CAR autoantigen in human cone cells. Differences between recoverin and visinin probably reflect species differences rather than rod-cone differences. Recoverin, or a nearly identical molecule, is present in mammalian cones and likely is the cone cell CAR autoantigen.
我们研究了一种假设,即视锥蛋白(visinin),一种最初在鸡视网膜中被鉴定的视锥细胞特异性蛋白,是恢复蛋白(recoverin)的视锥细胞同源物,并且可能是人类视锥细胞中癌症相关性视网膜病变(CAR)的自身抗原。
从鸡视网膜中纯化视锥蛋白,并检测其与CAR抗血清的结合情况。此外,视锥蛋白特异性抗体用于免疫细胞化学和蛋白质印迹分析,以确定视锥蛋白是否存在于人类或牛的视网膜中。针对恢复蛋白的抗肽抗体用于免疫细胞化学,以将恢复蛋白定位到哺乳动物的视锥细胞中。
CAR抗血清识别恢复蛋白而非视锥蛋白。此外,通过免疫细胞化学方法或尝试纯化该蛋白,在哺乳动物视网膜中均未检测到视锥蛋白。与视锥蛋白不同,针对恢复蛋白分子不同区域的特异性抗体可使人类视网膜中的视杆细胞和视锥细胞均染色。
视锥蛋白不是人类视锥细胞中的CAR自身抗原。恢复蛋白和视锥蛋白之间的差异可能反映了物种差异而非视杆细胞与视锥细胞的差异。恢复蛋白或一个几乎相同的分子存在于哺乳动物的视锥细胞中,并且可能是视锥细胞CAR自身抗原。