在大肠杆菌中表达的大鼠单胺氧化酶B含有共价结合的黄素腺嘌呤二核苷酸（FAD）。

Rat monoamine oxidase B expressed in Escherichia coli has a covalently-bound FAD.

作者信息

Hirashiki I, Ogata F, Ito A

机构信息

Department of Chemistry, Faculty of Science, Kyushu University, Fukuoka, Japan.

出版信息

Biochem Mol Biol Int. 1995 Sep;37(1):39-44.

PMID:8653086

Abstract

Rat liver monoamine oxidase B (MAO B) was expressed in E. coli as catalytically active form, though inclusion bodies of the enzyme were also formed as a major protein in the cell. The active form of the recombinant MAO B exhibited similar properties as rat liver enzyme and localized in membrane of the bacteria. Covalent attachment of FAD to polypeptide chain of the recombinant enzyme was revealed by a labeling experiment with [3H]-pargyline, an irreversible mechanism-based inhibitor, indicating that the covalent linkage of FAD to the apoprotein was formed even in the prokaryotic cell. This observation suggests autocatalytic formation of the linkage in MAO B.

摘要

大鼠肝脏单胺氧化酶B（MAO B）在大肠杆菌中以催化活性形式表达，不过该酶的包涵体也作为细胞中的主要蛋白质形成。重组MAO B的活性形式表现出与大鼠肝脏酶相似的特性，并定位于细菌膜中。用[3H]-帕吉林（一种基于不可逆机制的抑制剂）进行标记实验，揭示了FAD与重组酶多肽链的共价连接，这表明即使在原核细胞中，FAD与脱辅基蛋白的共价连接也会形成。这一观察结果表明MAO B中该连接是自动催化形成的。