MACROPHAGE OXIDATION OF L-ARGININE TO NITRITE AND NITRATE - NITRIC-OXIDE IS AN INTERMEDIATE
MACROPHAGE OXIDATION OF L-ARGININE TO NITRITE AND NITRATE - NITRIC-OXIDE IS AN INTERMEDIATE
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DOI：
10.1021/bi00424a003
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发表时间：
1988-11-29
期刊：
影响因子：
2.9
通讯作者：
WISHNOK, JS
中科院分区：
文献类型：
--
作者：
MARLETTA, MA;YOON, PS;WISHNOK, JS
先前的研究表明，用干扰素γ和大肠杆菌脂多糖免疫刺激的鼠巨噬细胞通过氧化两个化学等价的胍基氮中的一个，从L -精氨酸合成NO₂⁻、NO₃⁻和瓜氨酸。在从活化的RAW 264.7细胞分离的100000g上清液中发现了这种非常特殊反应的酶活性，而在未受刺激的细胞中完全不存在这种活性。这种活性需要NADPH和L -精氨酸，并且被Mg²⁺增强。当含有酶活性的亚细胞组分与L -精氨酸、NADPH和Mg²⁺一起孵育时，观察到一氧化氮的形成。一氧化氮的形成依赖于L -精氨酸和NADPH的存在，并且被NO₂⁻/NO₃⁻合成抑制剂NG -单甲基 - L -精氨酸抑制。此外，当与L -[胍基 - ¹⁵N₂]精氨酸一起孵育时，一氧化氮被¹⁵N标记。结果表明，一氧化氮是L -精氨酸转化为NO₂⁻ - NO₃⁻和瓜氨酸途径的中间产物。L -精氨酸是巨噬细胞激活到杀菌/杀肿瘤状态所必需的，这表明一氧化氮在这个激活过程中作为一种细胞内信号，其方式类似于最近在内皮细胞中观察到的情况，即一氧化氮导致血管平滑肌松弛[帕尔默，R.M.J.，阿什顿，D.S.，和蒙卡达，S.（1988年）《自然》（伦敦）333卷，664 - 666页]
Previous studies have shown that murine macrophages immunostimulated with interferon .gamma. and Escherichia coli lipopolysaccharide synthesize NO2-, NO3-, and citrulline from L-arginine by oxidation of one of the two chemically equivalent guanido nitrogens. The enzymatic activity for this very unusual reaction was found in the 100000g supernatant isolated from activated RAW 264.7 cells and was totally absent in unstimulated cells. This activity requires NADPH and L-arginine and is enhanced by Mg2+. When the subcellular fraction containing the enzyme activity was incubated with L-arginine, NADPH, and Mg2+, the formation of nitric oxide was observed. Nitric oxide formation was dependent on the presence of L-arginine and NADPH and was inhibited by the NO2-/NO3- synthesis inhibitor NG-monomethyl-L-arginine. Furthermore, when incubated with L-[guanido-15N2]arginine, the nitric oxide was 15N-labeled. The results show that nitric oxide is an intermediate in the L-arginine to NO2+- NO3-, and citrulline pathway. L-Arginine is required for the activation of macrophages to the bactericidal/tumoricidal state and suggests that nitric oxide is serving as an intracellular signal for this activation process in a manner similar to that very recently observed in endothelial cells, where nitric oxide leads to vascular smooth muscle relaxation [Palmer, R.M.J., Ashton, D.S., and Moncada, S. (1988) Nature (London) 333, 664-666].