Bailey J S, Siu C H
Banting and Best Department of Medical Research, University of Toronto, Ont. Canada.
Biochem Cell Biol. 1988 Jul;66(7):750-7. doi: 10.1139/o88-086.
A cellular retinoic acid-binding protein from 1-day-old mouse pups has been purified to homogeneity. The isolation procedure included gel filtration on Sephadex G-75, ion exchange chromatography on DEAE cellulose, and chromatofocusing on PBE9-4 ion exchange resin. The chromatofocusing step was most useful in removing the major contaminants, which were otherwise difficult to remove. The binding protein was finally subjected to two cycles of high performance liquid chromatography on a DEAE-5PW column to achieve homogeneity. The protein has an isoelectric point of 4.75 and consists of a single polypeptide, migrating with an apparent Mr of 14,600 in SDS--polyacrylamide gel electrophoresis. Amino-terminal sequence analysis showed that the mouse cellular retinoic acid-binding protein has a high percentage of amino acid identity with other retinoid-binding proteins. However, it is immunologically distinct from the cellular retinol-binding protein.
已将1日龄小鼠幼崽的一种细胞视黄酸结合蛋白纯化至同质。分离过程包括在Sephadex G - 75上进行凝胶过滤、在DEAE纤维素上进行离子交换色谱以及在PBE9 - 4离子交换树脂上进行色谱聚焦。色谱聚焦步骤对于去除主要污染物最为有用，否则这些污染物很难去除。最终，结合蛋白在DEAE - 5PW柱上进行了两个循环的高效液相色谱以实现同质。该蛋白的等电点为4.75，由一条单一多肽组成，在SDS - 聚丙烯酰胺凝胶电泳中表观Mr为14,600。氨基末端序列分析表明，小鼠细胞视黄酸结合蛋白与其他类视黄醇结合蛋白具有较高的氨基酸同一性百分比。然而，它在免疫上与细胞视黄醇结合蛋白不同。
