PURIFICATION OF SOLUBLE GUANYLATE CYCLASE FROM RAT-LIVER
PURIFICATION OF SOLUBLE GUANYLATE CYCLASE FROM RAT-LIVER
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DOI：
10.1073/pnas.76.1.219
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发表时间：
1979-01-01
影响因子：
11.1
通讯作者：
MURAD, F
中科院分区：
文献类型：
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作者：
BRAUGHLER, JM;MITTAL, CK;MURAD, F
从大鼠肝脏中提纯了可溶性鸟苷环化酶[GTP焦磷酸裂解酶(环化)，EC 4.6.1.2]，在聚丙烯酰胺凝胶上显示一条与活性一致的蛋白条带，其相对分子质量为150,000。以Mn2+为辅因子时，表观比活力为276nmolcGMP/mg/min，以Mg2+为辅因子时，表观比活力为23.8nmolcGMP/mg/min。这意味着Mn2+和Mg2+的活性分别纯化了9200倍和7400倍。可溶性鸟苷环化酶比活力不随蛋白质浓度变化而恒定。在纯化的所有阶段，增加鸟苷环化酶试验中的酶浓度都会增加制剂的表观比活力。纯化后的酶可被硝普钠、一氧化氮、花生四烯酸、亚油酸、油酸和超氧化物歧化酶激活。酶的活化程度依赖于所测酶蛋白的浓度。
Soluble guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] was purified from rat liver and exhibited a single protein band on polyacrylamide gels coincident with activity and indicative of MW of 150,000. The apparent specific activity of the purified enzyme was 276 nmol of cyclic[c]GMP formed/mg per min with Mn2+ as the cation of cofactor and 23.8 nmol of cGMP formed/mg per min with Mg2+. This represented 9200-fold and 7400-fold purifications of Mn2+ and Mg2+ activities, respectively. The specific activity of soluble guanylate cyclase was not constant with protein concentration. At all stages of purification, increasing the enzyme concentration in the guanylate cyclase assay increased the apparent specific activity of the preparation. The purified enzyme could be activiated by nitroprusside, nitric oxide, arachidonate, linoleate, oleate and superoxide dismutase. The degree of activation was dependent upon the concentration of enzyme protein assayed.