在大肠杆菌中作为谷胱甘肽-S-转移酶融合蛋白表达的人细胞视黄酸结合蛋白II的配体结合特性。
Ligand binding properties of human cellular retinoic acid binding protein II expressed in E. coli as a glutathione-S-transferase fusion protein.
作者信息
Redfern C P, Wilson K E
机构信息
Department of Medicine, Medical School, University of Newcastle, Newcastle upon Tyne, UK.
出版信息
FEBS Lett. 1993 Apr 26;321(2-3):163-8. doi: 10.1016/0014-5793(93)80100-9.
To test the hypothesis that 9-cis-retinoic acid is a ligand for cellular retinoic acid binding protein II (CRABP II), human CRABP II was expressed as a glutathione-S-transferase fusion protein (GST-CRABP II) and a single affinity purification step used to extract it from bacterial lysates. GST-CRABP II bound all trans-retinoic acid with high affinity (Kd 14.2 +/- 6.5 nM), but 9-cis-retinoic acid bound poorly. These studies suggest that 9-cis-retinoic acid is not a ligand for CRABP II. Their ease of purification makes GST-CRABP fusion proteins useful tools for ligand binding studies with different retinoids.
为了验证9-顺式视黄酸是细胞视黄酸结合蛋白II（CRABP II）的配体这一假设，将人CRABP II表达为谷胱甘肽-S-转移酶融合蛋白（GST-CRABP II），并采用单一亲和纯化步骤从细菌裂解物中提取该蛋白。GST-CRABP II与全反式视黄酸具有高亲和力结合（解离常数Kd为14.2±6.5 nM），但与9-顺式视黄酸结合较差。这些研究表明9-顺式视黄酸不是CRABP II的配体。其易于纯化的特性使GST-CRABP融合蛋白成为用于不同类视黄醇配体结合研究的有用工具。
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