ISOLATION OF NITRIC-OXIDE SYNTHETASE, A CALMODULIN-REQUIRING ENZYME
ISOLATION OF NITRIC-OXIDE SYNTHETASE, A CALMODULIN-REQUIRING ENZYME
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DOI：
10.1073/pnas.87.2.682
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发表时间：
1990-01-01
影响因子：
11.1
通讯作者：
SNYDER, SH
中科院分区：
文献类型：
--
作者：
BREDT, DS;SNYDER, SH
一氧化氮介导内皮细胞的血管松弛作用、巨噬细胞和中性粒细胞的细胞毒作用以及兴奋性氨基酸对小脑cGMP的影响。由一种与化学计量比生产瓜氨酸相关的可溶酶从精氨酸中酶解形成瓜氨酸需要NADPH和钙离子。我们发现一氧化氮合酶的活性需要钙调素。利用NADPH洗脱的2‘’，5‘-ADP亲和层析柱，我们从大鼠小脑中纯化了6000倍的一氧化氮合酶。纯化后的酶在SDS/PAGE上呈150 kDa的单一条带迁移，天然酶为单体。
Nitric oxide mediates vascular relaxing effects of endothelial cells, cytotoxic actions of macrophages and neutrophils, and influences of excitatory amino acids on cerebellar cyclic GMP. Its enzymatic formation from arginine by a soluble enzyme associated with stoichiometric production of citrulline requires NADPH and Ca2+. We show that nitric oxide synthetase activity requires calmodulin. Utilizing a 2'',5''-ADP affinity column eluted with NADPH, we have purified nitric oxide synthetase 6000-fold to homogeneity from rat cerebellum. The purified enzyme migrates as a single 150-kDa band on SDS/PAGE, and the native enzyme appears to be a monomer.