CLONING AND CHARACTERIZATION OF INDUCIBLE NITRIC-OXIDE SYNTHASE FROM MOUSE MACROPHAGES
CLONING AND CHARACTERIZATION OF INDUCIBLE NITRIC-OXIDE SYNTHASE FROM MOUSE MACROPHAGES
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DOI：
10.1126/science.1373522
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发表时间：
1992-04-10
期刊：
影响因子：
56.9
通讯作者：
NATHAN, C
中科院分区：
文献类型：
--
作者：
XIE, QW;CHO, HJ;NATHAN, C
一氧化氮（NO）在细胞间传递多种信息，包括血管舒张、神经传递和细胞毒性信号。在一些内皮细胞和神经元中，组成型NO合酶被升高细胞内钙浓度并促进钙调蛋白结合的激动剂瞬时激活。相反，在巨噬细胞中，NO合酶活性在细胞暴露于细胞因子和细菌产物后缓慢出现，持续存在，并且独立于钙和钙调蛋白发挥作用。一种单特异性抗体被用来克隆互补DNA，编码两种亚型的NO合酶从免疫激活的小鼠巨噬细胞。采用液相色谱-质谱联用技术对大部分氨基酸序列进行确认。巨噬细胞NO合酶与小脑NO合酶有很大不同。巨噬细胞酶在转录水平上被免疫诱导，并且非常类似于经苦参碱处理的肿瘤细胞和炎性中性粒细胞中的酶。
Nitric oxide (NO) conveys a variety of messages between cells, including signals for vasorelaxation, neurotransmission, and cytotoxicity. In some endothelial cells and neurons, a constitutive NO synthase is activated transiently by agonists that elevate intracellular calcium concentrations and promote the binding of calmodulin. In contrast, in macrophages, NO synthase activity appears slowly after exposure of the cells to cytokines and bacterial products, is sustained, and functions independently of calcium and calmodulin. A monospecific antibody was used to clone complementary DNA that encoded two isoforms of NO synthase from immunologically activated mouse macrophages. Liquid chromatography-mass spectrometry was used to confirm most of the amino acid sequence. Macrophage NO synthase differs extensively from cerebellar NO synthase. The macrophage enzyme is immunologically induced at the transcriptional level and closely resembles the enzyme in cytokine-treated tumor cells and inflammatory neutrophils.