Improved purification of brine-shrimp (Artemia saline) (Na+ + K+)-activated adenosine triphosphatase and amino-acid and carbohydrate analyses of the isolated subunits.
Improved purification of brine-shrimp (Artemia saline) (Na+ + K+)-activated adenosine triphosphatase and amino-acid and carbohydrate analyses of the isolated subunits.
复制标题
改进丰年虾（Artemia saline）（Na K ）激活的三磷酸腺苷酶的纯化以及分离亚基的氨基酸和碳水化合物分析。
DOI：
10.1042/bj1920107
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发表时间：
1980
期刊：
影响因子：
--
通讯作者：
Hokin,LE
中科院分区：
文献类型：
--
作者：
Peterson,GL;Hokin,LE
通过使用Lubrol WX和非变性浓度的十二烷基硫酸钠（SDS），（Na+ + K+）活化的ATP酶的纯化在纯度和产率方面都比以前的方法[Peterson，Ewing，Hootman & Conte（1978）J.Biol.Chem.253，4762-4770]提高了2倍。该酶纯化200倍以上的匀浆。该制剂具有约600 μ mol Pi/h/mg蛋白质的比活性，并且根据考马斯亮蓝染色的SDS/聚丙烯酰胺凝胶的定量，纯度约为60%。每100 g卤虫卵中蛋白质含量约为10 mg。该方法非常适合于小规模（10- 25 g干孢囊）或大规模（900 g干孢囊）的纯化，并且描述了用于两者的方法。在Bio-Gel A 1.5m上通过SDS-凝胶过滤以纯形式分离大的（Na+ + K+）-活化的ATP酶亚基（α-亚基）。在Bio-Gel柱上用其他污染蛋白洗脱小亚基（β-亚基），但通过从SDS/聚丙烯酰胺凝胶中提取以纯形式分离。两个亚基的氨基酸和碳水化合物组成的报告。α-亚基含有5.2%重量的碳水化合物，β-亚基含有9.2%重量的碳水化合物。唾液酸是从两个亚基缺席。
Purification of the (Na+ + K+)-activated ATPase has been improved 2-fold the respect to both purity and yield over the previous method [Peterson, Ewing, Hootman & Conte (1978) J. Biol. Chem. 253, 4762-4770] by using Lubrol WX and non-denaturing concentrations of sodium dodecyl sulphate (SDS). The enzyme was purified 200-fold over the homogenate. The preparation had a specific activity of about 600 mumol of Pi/h per mg of protein, and was about 60% pure according to quantification of Coomassie Blue-stained SDS/polyacrylamide gels. The yield of purified enzyme was about 10 mg of protein per 100g of dry brine-shrimp (Artemia salina) cysts. The method is highly suitable for purification either on a small scale (10-25g of dry cysts) or on a large scale (900g of dry cysts) and methods are described for both. The large (Na+ + K+)-activated ATPase subunit (alpha-subunit) was isolated in pure form by SDS-gel filtration on Bio-Gel A 1.5m. The small subunit (beta-subunit) was eluted with other contaminating proteins on the Bio-Gel column, but was isolated in pure form by extraction from SDS/polyacrylamide gels. The amino acid and carbohydrate compositions of both subunits are reported. The alpha-subunit contained 5.2% carbohydrate by weight, and the beta-subunit 9.2%. Sialic acid was absent from both subunits.