Schmidt H H, Murad F
Northwestern University Medical School, Dept. of Pharmacology, Chicago, IL 60611.
Biochem Biophys Res Commun. 1991 Dec 31;181(3):1372-7. doi: 10.1016/0006-291x(91)92090-7.
A NO synthase (NOS, EC 1.14.23) was isolated from human cerebellum by two sequential chromatography steps, that is affinity chromatography on 2'5'ADP sepharose and size exclusion chromatography on Superose 6. Human NOS migrated as a single band of 160 kDa on SDS/PAGE. The enzyme was Ca2+/calmodulin-regulated and NADPH/tetrahydrobiopterin (BH4)-dependent, which are characteristics of a type I NOS previously isolated from rat cerebellum. Antisera raised against purified rat cerebellar NOS crossreacted specifically with a 160 kDa protein in crude supernatant fraction of human cerebellum and purified human NOS but not in crude supernatant fraction of the temporal lobe. These findings provide evidence that nitrinergic signal transduction through conversion of L-arginine to L-citrulline and NO does also occur in humans and NO may function as a neurotransmitter in the human central nervous system.
通过两步连续色谱法从人小脑中分离出一种一氧化氮合酶（NOS，EC 1.14.23），即2',5'-ADP琼脂糖亲和色谱和Superose 6分子筛色谱。人NOS在SDS/PAGE上迁移为一条160 kDa的单带。该酶受Ca2+/钙调蛋白调节，依赖NADPH/四氢生物蝶呤（BH4），这是先前从大鼠小脑中分离出的I型NOS的特征。针对纯化的大鼠小脑NOS产生的抗血清与人小脑粗上清液组分和纯化的人NOS中的160 kDa蛋白特异性交叉反应，但与颞叶粗上清液组分不发生交叉反应。这些发现提供了证据，表明通过将L-精氨酸转化为L-瓜氨酸和NO的硝能信号转导在人类中也会发生，并且NO可能在人类中枢神经系统中作为神经递质发挥作用。
