从牛视网膜和视网膜色素上皮中纯化细胞视黄醛结合蛋白。

Purification of cellular retinaldehyde-binding protein from bovine retina and retinal pigment epithelium.

作者信息

Saari J C, Bredberg D L

机构信息

Department of Ophthalmology, University of Washington, School of Medicine, Seattle 98195.

出版信息

Exp Eye Res. 1988 Apr;46(4):569-78. doi: 10.1016/s0014-4835(88)80013-7.

DOI:10.1016/s0014-4835(88)80013-7

PMID:2838311

Abstract

Cellular retinaldehyde-binding protein (CRALP) has been purified from extracts of bovine retina or retinal pigment epithelium by a procedure employing an initial, high-capacity anion exchange chromatographic step and anion exchange HPLC for removal of a persistent contaminant. The procedure also yields fractions containing three other retinoid-binding proteins present in retina (cellular retinol-, cellular retinoic acid- and interphotoreceptor retinol-binding proteins; CRBP, CRABP and IRBP, respectively). Procedures are described for labeling CRALBP with 9-cis-retinaldehyde, 11-cis-retinaldehyde, or 11-cis-retinol. There are approx. 3 nmol of CRALBP per adult bovine eye and the binding protein is ca. 0.5% of the soluble protein of a retinal supernatant.

摘要

细胞视黄醛结合蛋白（CRALP）已通过以下步骤从牛视网膜或视网膜色素上皮提取物中纯化得到：首先采用高容量阴离子交换色谱步骤，然后用阴离子交换高效液相色谱法去除一种持久性污染物。该方法还能得到含有视网膜中存在的其他三种类视黄醇结合蛋白的组分（分别为细胞视黄醇结合蛋白、细胞视黄酸结合蛋白和光感受器间视黄醇结合蛋白；CRBP、CRABP和IRBP）。文中描述了用9-顺式视黄醛、11-顺式视黄醛或11-顺式视黄醇标记CRALBP的方法。每只成年牛眼约有3 nmol的CRALBP，该结合蛋白约占视网膜上清液可溶性蛋白的0.5%。