细菌表达的大鼠视黄醇结合蛋白对视黄醇和转甲状腺素蛋白具有结合功能。

Bacterially expressed rat retinol-binding protein is functional for retinol and transthyretin bindings.

作者信息

Yamamoto Y, Yoshizawa T, Mano H, Masushige S, Kato S

机构信息

Department of Agricultural Chemistry, Faculty of Agriculture, Tokyo University of Agriculture, Japan.

出版信息

J Nutr Sci Vitaminol (Tokyo). 1996 Aug;42(4):257-66. doi: 10.3177/jnsv.42.257.

DOI:10.3177/jnsv.42.257

PMID:8906627

Abstract

Retinol-binding protein (RBP) was expressed in Escherichia coli using the cDNA for rat RBP, and characterized. The expressed RBP was fused to maltose-binding protein (MBP) at the N-terminal end (MBP-RBP), and MBP was enzymatically removed from the MBP-RBP with proteinase factor Xa. The binding of retinol and transthyretin (TTR) to the recombinant RBP was monitored by means of gel filtration. The recombinant RBP specifically bound to retinol with an affinity similar to that of purified RBP from rat serum. Furthermore, the retinol-bound recombinant RBP formed hetero-complexes with TTR similar to RBP. Thus, the results showed that the recombinant RBP expressed in E. coli is as functional as serum RBP in terms of retinol and TTR bindings.

摘要

利用大鼠视黄醇结合蛋白（RBP）的cDNA在大肠杆菌中表达并对其进行了表征。所表达的RBP在N端与麦芽糖结合蛋白（MBP）融合（MBP-RBP），然后用蛋白酶因子Xa从MBP-RBP中酶解去除MBP。通过凝胶过滤监测视黄醇和转甲状腺素蛋白（TTR）与重组RBP的结合。重组RBP与视黄醇特异性结合，其亲和力与大鼠血清纯化的RBP相似。此外，结合视黄醇的重组RBP与TTR形成异源复合物，类似于RBP。因此，结果表明在大肠杆菌中表达的重组RBP在视黄醇和TTR结合方面与血清RBP具有相同的功能。