Monoclonal antibody modification of the ATPase activity of Escherichia coli F1 ATPase.
Monoclonal antibody modification of the ATPase activity of Escherichia coli F1 ATPase.
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单克隆抗体修饰大肠杆菌 F1 ATP 酶的 ATP 酶活性。
DOI：
10.1021/bi00497a013
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发表时间：
1990
期刊：
影响因子：
2.9
通讯作者：
R. Capaldi
中科院分区：
文献类型：
--
作者：
R. Aggeler;J. Mendel;R. Capaldi
针对ECF1的五个亚基(α、β、γ、β和epsilon)中的每一个都制备了单抗，这些抗体被用于拓扑学研究和单个亚基在酶功能中的作用的研究。所获得的单抗均与ECF1反应，但有几个与ECF1F0反应失败，包括3个抗γ亚基的mAb(Gamma II、Gamma III和Gamma IV)，1个抗Delta的mAb，以及2个抗epsilon的mAb(epsilon I和epsilon II)。这些拓扑数据与位于复合体F1和F0部分之间的界面上的Gamma、Delta和epsilon亚基一致。用两种形式的ECF1来研究单抗对酶的ATPase活性的影响：ECF1与epsilon亚基紧密结合并起抑制活性的作用；ECF1*通过有机溶剂处理去除了Delta和epsilon亚基。ECF1*在标准条件下具有ATPase活性，其标准条件为93摩尔的ATP水解物min-1 mg-1，cf。我们的标准ECF1制剂的酶活为7.5U mg-1，经胰酶处理去除了epsilon亚基的酶的活性为mg-1。在一个复杂的过程中，ECF1*的蛋白酶将活性降低到单位mg-1，该过程包括通过切割α亚基来抑制活性，通过切割伽玛亚基来激活活性，通过切割β亚单位来抑制活性。密度梯度离心实验证明，通过改变酶与epsilon亚单位的亲和力，针对Gamma亚基的单抗Gamma II和Gamma III分别激活ECF1 4.4倍和2.4倍。γ-亚基单抗不改变ECF1*或胰酶处理的酶的ATPase活性。α亚基单抗对ECF1的激活倍数为2.5倍，对ECF1F0的激活倍数为1.3倍，但对ECF1*的ATPase活性抑制30%。
Monoclonal antibodies (mAbs) have been made against each of the five subunits of ECF1 (alpha, beta, gamma, delta, and epsilon), and these have been used in topology studies and for examination of the role of individual subunits in the functioning of the enzyme. All of the mAbs obtained reacted with ECF1, while several failed to react with ECF1F0, including three mAbs against the gamma subunit (gamma II, gamma III, and gamma IV), one mAb against delta, and two mAbs against epsilon (epsilon I and epsilon II). These topology data are consistent with the gamma, delta, and epsilon subunits being located at the interface between the F1 and F0 parts of the complex. Two forms of ECF1 were used to study the effects of mAbs on the ATPase activity of the enzyme: ECF1 with the epsilon subunit tightly bound and acting to inhibit activity and ECF1* in which the delta and epsilon subunits had been removed by organic solvent treatment. ECF1* had an ATPase activity under standard conditions of 93 mumol of ATP hydrolyzed min-1 mg-1, cf. an activity of 7.5 units mg-1 for our standard ECF1 preparation and 64 units mg-1 for enzyme in which the epsilon subunit had been removed by trypsin treatment. The protease digestion of ECF1* reduced activity to 64 units mg-1 in a complicated process involving an inhibition of activity by cleavage of the alpha subunit, activation by cleavage of gamma, and inhibition with cleavage of the beta subunit. mAbs to the gamma subunit, gamma II and gamma III, activated ECF1 by 4.4- and 2.4-fold, respectively, by changing the affinity of the enzyme for the epsilon subunit, as evidenced by density gradient centrifugation experiments. The gamma-subunit mAbs did not alter the ATPase activity of ECF1*- or trypsin-treated enzyme. The alpha-subunit mAb (alpha I) activated ECF1 by a factor of 2.5-fold and ECF1F0 by 1.3-fold, but inhibited the ATPase activity of ECF1* by 30%.
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DOI：
--
发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Richter,ML;Patrie,WJ;McCarty,RE
通讯作者：
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影响因子：
2.9
作者：
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通讯作者：
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从脂质双层的头基区域标记大肠杆菌的 ATP 合酶。
影响因子：
2.9
作者：
Aggeler,R;Zhang,YZ;Capaldi,RA
通讯作者：
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影响因子：
2.9
作者：
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DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Dunn,SD
通讯作者：
Dunn,SD