Oprian D D, Asenjo A B, Lee N, Pelletier S L
Graduate Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02254.
Biochemistry. 1991 Dec 3;30(48):11367-72. doi: 10.1021/bi00112a002.
Color vision in humans is mediated by three pigments from retinal cone photoreceptor cells: blue, green, and red. We have designed and chemically synthesized genes for each of these three pigments. The genes were expressed in COS cells, reconstituted with 11-cis-retinal chromophore, and purified to homogeneity using an immunoaffinity procedure. To facilitate the immunoaffinity purification, each pigment was modified at the carboxy terminus to contain an additional eight amino acid epitope for a monoclonal antibody previously used to purify bovine rhodopsin. The spectra for the isolated pigments had maxima of 424, 530, and 560 nm, respectively, for the blue, green, and red pigments. These maxima are in excellent agreement with the maxima previously observed by microspectrophotometry of individual human cone cells. The spectra are the first to be obtained from isolated human color vision pigments. They confirm the original identification of the three color vision genes, which was based on genetic evidence [Nathans, J., Thomas, D., & Hogness, D.S. (1986) Science 232, 193].
蓝色、绿色和红色。我们已经为这三种色素中的每一种设计并化学合成了基因。这些基因在COS细胞中表达，与11-顺式视黄醛发色团重组，并使用免疫亲和程序纯化至同质。为便于免疫亲和纯化，每种色素在羧基末端进行了修饰，以包含一个额外的八氨基酸表位，用于先前用于纯化牛视紫红质的单克隆抗体。分离出的色素的光谱中，蓝色、绿色和红色色素的最大值分别为424、530和560纳米。这些最大值与之前通过对单个人类视锥细胞进行显微分光光度法观察到的最大值非常吻合。这些光谱是首次从分离出的人类色觉色素中获得的。它们证实了基于遗传证据对三种色觉基因的最初鉴定【内森斯，J.，托马斯，D.，&霍格尼斯，D.S.（1986年）《科学》232，193】。
