Expression of recombinant myeloperoxidase using a baculovirus expression system.
Expression of recombinant myeloperoxidase using a baculovirus expression system.
复制标题
使用杆状病毒表达系统表达重组髓过氧化物酶。
DOI：
10.1016/0006-291x(92)90482-z
复制
发表时间：
1992
影响因子：
3.1
通讯作者：
KinkadeJr,JM
中科院分区：
文献类型：
--
作者：
Taylor,KL;Uhlinger,DJ;KinkadeJr,JM
髓过氧化物酶（MPO）是一种存在于正常人中性粒细胞嗜天青颗粒中的含血红素的糖基化酶。这种酶通过催化强氧化剂次氯酸的形成在宿主防御系统的杀微生物活性中起主要作用。虽然MPO的氨基酸序列已从cDNA中推导出来，但这种酶所观察到的异质性的结构基础尚不清楚。此外，辅基的性质及其与脱辅基蛋白的连接方式尚未确定。为了解决有关MPO的结构特征，这是在复杂的翻译后加工过程中出现的这种酶的问题，我们利用杆状病毒系统在Sf9昆虫细胞中表达MPO。观察到两种糖基化的MPO单链前体物质：分泌的84 kDa物质和细胞相关的74 kDa物质。这是第一次报告的表达系统中，细胞相关的MPO前体进行翻译后蛋白水解加工。
Myeloperoxidase (MPO) is a glycosylated heme-containing enzyme present in the azurophilic granules of normal human polymorphonuclear neutrophils. This enzyme plays a major role in the microbicidal activity of the host defense system by catalyzing the formation of the potent oxidant, hypochlorous acid. Although the amino acid sequence of MPO has been deduced from the cDNA, the structural basis for the observed heterogeneity of this enzyme is not known. Furthermore, the nature of the prosthetic group and its mode of linkage to the apoprotein has not been determined. To address questions regarding the structural features of MPO, which arise during the complex posttranslational processing of this enzyme, we utilized a baculovirus system to express MPO in Sf9 insect cells. Two glycosylated, single-chain precursor species of MPO were observed: an 84 kDa species that was secreted and a 74 kDa species that was cell-associated. This is the first report of an expression system in which a cell-associated MPO precursor undergoes posttranslational proteolytic processing.