Barroso J B, Corpas F J, Carreras A, Sandalio L M, Valderrama R, Palma J M, Lupiáñez J A, del Río L A
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Experimentales, Universidad de Jaén, Paraje "Las Lagunillas" s/n, E-23071 Jaén, Spain.
J Biol Chem. 1999 Dec 17;274(51):36729-33. doi: 10.1074/jbc.274.51.36729.
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 Ca(2+)-dependent NOS activity of 5.61 nmol of L-[(3)H]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 immunogold-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.
对豌豆（Pisum sativum L.）叶片过氧化物酶体中一氧化氮合酶（NOS）的存在情况进行了研究。通过差速离心和蔗糖密度梯度离心法对植物细胞器进行了纯化。在纯化的完整过氧化物酶体中，测得Ca(2+)依赖性NOS活性为5.61 nmol L-[(3)H]瓜氨酸mg(-1)蛋白min(-1)，而线粒体中未检测到活性。过氧化物酶体的NOS活性受到哺乳动物NO合酶不同特征明确的抑制剂的明显抑制（60 - 90%）。用针对小鼠诱导型NOS C末端区域的多克隆抗体对过氧化物酶体进行免疫印迹分析，显示出一条130 kDa的免疫反应性蛋白条带。电子显微镜免疫金标记证实了NOS在过氧化物酶体基质以及叶绿体中的亚细胞定位。过氧化物酶体中存在NOS表明这些氧化细胞器是细胞内一氧化氮（NO）的来源，并暗示了过氧化物酶体在细胞信号转导机制中的新作用。
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