Localization of nitric-oxide synthase in plant peroxisomes
Localization of nitric-oxide synthase in plant peroxisomes
复制标题
DOI：
10.1074/jbc.274.51.36729
复制
发表时间：
1999-12-17
影响因子：
4.8
通讯作者：
del Río, LA
中科院分区：
文献类型：
--
作者：
Barroso, JB;Corpas, FJ;del Río, LA
豌豆（PisumsativumL.）研究了用差速离心和蔗糖密度梯度离心纯化植物细胞器。在纯化的完整过氧化物酶体中，测得5.61 nmol L-[H-3]瓜氨酸mg（-1）蛋白/min（-1）的Ca 2+依赖性NOS活性，而在线粒体中未检测到活性。过氧化物酶体NOS活性明显抑制（60-90%）由不同的哺乳动物NO脱氢酶的良好表征的抑制剂。过氧化物酶体的免疫印迹分析与多克隆抗体对小鼠诱导型一氧化氮合酶的C末端区域显示了130 kDa的免疫反应性蛋白。电子显微镜免疫金标记证实了NOS在过氧化物酶体基质和叶绿体中的亚细胞定位。NOS在过氧化物酶体中的存在表明这些氧化细胞器是一氧化氮（NO）的细胞来源，并暗示过氧化物酶体在细胞信号转导机制中的新作用。
The presence of nitric-oxide synthase (NOS) in peroxisomes from leaves of pea plants (Pisum sativum L.) was studied. Plant organelles were purified by differential and sucrose density gradient centrifugation. In purified intact peroxisomes a Ca2+-dependent NOS activity of 5.61 nmol of L-[H-3]citrulline mg(-1) protein min(-1) was measured while no activity was detected in mitochondria. The peroxisomal NOS activity was clearly inhibited (60-90%) by different well characterized inhibitors of mammalian NO synthases. The immunoblot analysis of peroxisomes with a polyclonal antibody against the C terminus region of murine iNOS revealed an immunoreactive protein of 130 kDa. Electron microscopy immuno-gold-labeling confirmed the subcellular localization of NOS in the matrix of peroxisomes as well as in chloroplasts. The presence of NOS in peroxisomes suggests that these oxidative organelles are a cellular source of nitric oxide (NO) and implies new roles for peroxisomes in the cellular signal transduction mechanisms.