人5-脂氧合酶cDNA在大肠杆菌中的表达。

Expression of human 5-lipoxygenase cDNA in Escherichia coli.

作者信息

Noguchi M, Matsumoto T, Nakamura M, Noma M

机构信息

Life Science Research Laboratory, Japan Tobacco Inc., Yokohama.

出版信息

FEBS Lett. 1989 Jun 5;249(2):267-70. doi: 10.1016/0014-5793(89)80638-6.

DOI:10.1016/0014-5793(89)80638-6

PMID:2661265

Abstract

A cDNA for human 5-lipoxygenase (5LO) was inserted into the vector pKC (constructed from pKK223-3 by replacing its replication origin with that of pUC18) and expressed in Escherichia coli. The enzyme expressed was purified to homogeneity from the cellular soluble fraction. The purified enzyme showed both 5LO and leukotriene A4 synthase activities, which were stimulated by Ca2+ and ATP. Its molecular mass (78 kDa) and NH2-terminal sequence were identical with those of 5LO purified from human leukocytes. The availability of the expression system will facilitate further studies on its regulation and the reaction mechanism of the enzyme.

摘要

将人5-脂氧合酶（5LO）的互补DNA（cDNA）插入载体pKC（由pKK223-3构建而成，通过用pUC18的复制起点替换其复制起点），并在大肠杆菌中表达。表达的酶从细胞可溶性部分纯化至同质。纯化的酶显示出5LO和白三烯A4合酶活性，这两种活性均受Ca2+和ATP刺激。其分子量（78 kDa）和氨基末端序列与人白细胞中纯化的5LO相同。该表达系统的可用性将有助于对其调节和酶的反应机制进行进一步研究。