PURIFICATION AND CDNA SEQUENCE OF AN INDUCIBLE NITRIC-OXIDE SYNTHASE FROM A HUMAN TUMOR-CELL LINE
PURIFICATION AND CDNA SEQUENCE OF AN INDUCIBLE NITRIC-OXIDE SYNTHASE FROM A HUMAN TUMOR-CELL LINE
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DOI：
10.1021/bi00094a017
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发表时间：
1993-11-02
期刊：
影响因子：
2.9
通讯作者：
WOOD, ER
中科院分区：
文献类型：
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作者：
SHERMAN, PA;LAUBACH, VE;WOOD, ER
多种细胞因子联合诱导一氧化氮合酶（NOS）在人结直肠癌细胞系DLD-1中的表达。我们纯化了这种酶，并检测了它的一些生化特性。小鼠巨噬细胞诱导的NOS的抗血清与DLD-1 NOS交叉反应。纯化的人和小鼠酶显示出类似的对外源钙和钙调素缺乏依赖的活性，这与纯化的脑和内皮型NOS对钙和钙调素的需求形成对比。我们还从DLD-1细胞中分离了细胞因子诱导的NOS的cDNA。对该cDNA和来自人肝脏、平滑肌和巨噬细胞的NOS cDNA的序列分析表明，在遗传水平上，人诱导的NOS存在单一的同种异构体。
A combination of cytokines induced the expression of nitric oxide synthase (NOS) in a human colorectal adenocarcinoma cell line, DLD-1. We have purified the enzyme and examined some of its biochemical properties. An antiserum to an inducible NOS from murine macrophages cross-reacted with the DLD-1 NOS. The purified human and murine enzymes displayed a similar lack of dependence on exogenous calcium and calmodulin for activity, which contrasts with the requirement for calcium and calmodulin of purified brain and endothelial isoforms of NOS. We have also isolated a cDNA for a cytokine-induced NOS from DLD-1 cells. Sequence analysis of this cDNA and NOS cDNAs from human liver, smooth muscle, and macrophages suggests that, at the genetic level, there is a single isoform of human-inducible NOS.