Reconstitution of adenosine triphosphatase of thermophilic bacterium from purified individual subunits.
Reconstitution of adenosine triphosphatase of thermophilic bacterium from purified individual subunits.
复制标题
从纯化的单个亚基中重建嗜热细菌的腺苷三磷酸酶。
DOI：
10.1016/s0021-9258(17)40416-9
复制
发表时间：
1977
期刊：
影响因子：
--
通讯作者：
Y. Kagawa
中科院分区：
文献类型：
--
作者：
M. Yoshida;N. Sone;H. Hirata;Y. Kagawa
1.在8 M尿素存在下，通过离子交换色谱法纯化来自嗜热细菌PS3的ATP酶的五个亚基（α、β、γ、δ和β-ATP酶）。然后，通过混合亚基溶液并在20-45度，pH 6.3至7.0下孵育它们来重建ATP酶活性。2.含有β + γ或α + β + δ的混合物恢复了ATP水解活性，但α + β和β + δ的混合物没有。不包括β的组合均无活性。3.从α + β + δ重建的ATP酶活性是热不稳定的，对NaN 3不敏感，而从含有β和γ的混合物获得的活性是热稳定的，对NaN 3敏感，像天然ATP酶一样。4.含有β和γ亚基的组件在凝胶电泳上具有与天然ATP酶分子相同的迁移率，那些没有γ亚基的组件更快地向阳极移动。5.亚基α和δ不抑制组装体（α + β + γ）或天然ATP酶的ATP酶活性。
1. Five subunits (alpha, beta, gamma, delta, and epsilon) of an ATPase from a thermophilic bacterium PS3 were purified in the presence of 8 M urea by ion exchange chromatography. Then the ATPase activity was reconstituted by mixing the subunit solutions and incubating them at 20-45 degrees, at pH 6.3 to 7.0. 2. Mixtures containing beta + gamma or alpha + beta + delta regained ATP-hydrolyzing activity, but mixtures of alpha + beta and beta + delta did not. Combinations not including beta were all inactive. 3. The ATPase activity reconstituted from alpha + beta + delta was thermolabile and insensitive to NaN3, whereas the activities obtained from mixtures containing beta and gamma were thermostable and sensitive to NaN3, like the native ATPase. 4. The assemblies containing both beta and gamma subunits had the same mobility as the native ATPase molecule on gel electrophoresis, those without the gamma subunit moved more rapidly toward the anode. 5. Subunits epsilon and delta did not inhibit the ATPase activity of either the assembly (alpha + beta + gamma) or the native ATPase.