摘要
目的纯化大鼠小脑一氧化氮合酶并进行特性研究。方法采用改良的DE52及2′,5′-ADP-agarose二步层析法。结果①大鼠小脑组织一氧化氮合酶(NOS)纯化6000倍,收率30%,SDS-PAGE成一条区带,分子量约为150ku;②纯化后的NOS活性依赖Ca2+和CaM,其半激活浓度分别为200nmol/L和10nmol/L,并可被TFP和NO2-Arg抑制,其半抑制浓度分别为10μmol/L和0.6μmol/L。结论NOS经改良后的二步层析法纯化所得结果与文献报道结果符合,此方法可用于进一步的实验研究。
Objective To purify NOS from rat cerebellum and study the character.Methods Cytosolic nitric oxide synthase(NOS)from rat cerebellum was purified to hemogenity by improved sequential chromotographa steps on DE52 and 2′,5′-ADP-agarose.Results NOS was purified by 6000 fold,and migrated as a single band on SDS-PAGE.the molecular weight was about 150ku.the activity was dependent on Ca2+/CaM,and was inhibited by TFP and NO2-Arg.Conclusions The results from this improved two sequential chromotographa steps are consistant from reports.So this improved method may be practical for further experiments.
出处
《徐州医学院学报》
CAS
1998年第2期93-96,共4页
Acta Academiae Medicinae Xuzhou
关键词
一氧化氮合酶
纯化
小脑
大鼠
Nitric oxide synthase Purification Cerebellum Rat