Synthetic inhibitors of adenylate kinases in the assays for ATPases and phosphokinases.
Synthetic inhibitors of adenylate kinases in the assays for ATPases and phosphokinases.
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ATP 酶和磷酸激酶测定中的腺苷酸激酶合成抑制剂。
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发表时间：
1975
期刊：
影响因子：
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通讯作者：
R. Schirmer
中科院分区：
文献类型：
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作者：
P. Feldhau;T. Fröhlich;R. Goody;M. Isakov;R. Schirmer
1. 程序给出了合成α， ω -二核苷5'-多磷酸盐作为腺苷酸激酶抑制剂。观察到抑制猪肌腺苷酸激酶的能力顺序为：Ap5A大于1；N6-etheno-Ap5A大于Ap6A大于Gp5A大于Ap4A大于Up5A。介绍了Ap5A的原料四磷酸腺苷的合成过程。2. 一个猪肌腺苷酸激酶分子与一个Ap5A分子结合。ap5a -腺苷酸激酶在271 nm处的差谱（最大值为5050 M-1 - cm-1）以及1:n6 -乙烯- ap5a的荧光性质可用于动力学和结合研究。3. 利用带负电荷的Ap5A特异性结合制备人肌腺苷酸激酶。酶被纯化至均匀性，总产率为65%，绝对值为70 mg / kg肌肉。4. 检测Ap5A对不同细胞及细胞器提取物中腺苷酸激酶的影响。Ap5A与其他核苷酸的比例为1:50 (mol/mol)，可抑制哺乳动物和昆虫骨骼肌、人红细胞和金黄色葡萄球菌提取物的腺苷酸激酶活性。烟叶和菠菜叶绿体中抑制腺苷酸激酶所需的比例为1:5，牛肝线粒体、人肾匀浆和大肠杆菌中抑制腺苷酸激酶所需的比例为2:1。Ap5A似乎不会在上述任何提取物中代谢。这些结果表明，Ap5A可以用于评估腺苷酸激酶在能量转导系统中从ADP产生ATP的贡献。5. 污染腺苷酸激酶可以被浓度的Ap5A抑制，而Ap5A不会干扰许多（磷酸）激酶和atp酶的研究。Ap5A在核苷二磷酸激酶测定和收缩蛋白力学生化特性研究中的应用是代表性的例子。Ap5A的使用使得研究ADP本身在这类系统中的作用成为可能。6. sepharose结合Ap5A用于从肌球蛋白和肌酸激酶样品中去除腺苷酸激酶的痕迹。7. 在Ap5A存在的情况下，测定静脉血溶血血清、毛细血管血浆和诱导完全溶血后全血样品中肌酸激酶的活性。这些发现在心肌梗死和肌营养不良的病例中具有临床意义。
1. Procedures are given for the syntheses of alpha,omega-dinucleoside 5'-polyphosphates as inhibitors of adenylate kinases. The following order for the ability of inhibiting pig muscle adenylate kinase was observed: Ap5A greater than 1:N6-etheno-Ap5A greater than Ap6A greater than Gp5A greater than Ap4A greater than Up5A. The synthesis of adenosine tetraphosphate, the starting material for Ap5A, is also described. 2. One molecule of pig muscle adenylate kinase binds one molecule of Ap5A. The difference spectrum of Ap5A-adenylate kinase with its maximum of 5050 M-1 - cm-1 at 271 nm, as well as the fluorescence properties of 1:N6-etheno-Ap5A can be used for kinetic and binding studies. 3. The specific binding of the negatively charged Ap5A was exploited in the preparation of human muscle adenylate kinase. The enzyme was purified to homogeneity with an overall yield of 65%, the absolute value being 70 mg per kg of muscle. 4. The effect of Ap5A on adenylate kinase in extracts of various cells and cell organelles was tested. A ratio of 1:50 (mol/mol) for Ap5A to other nucleotides was used for suppressing the adenylate kinase activity in extracts of mammalian and insect skeletal muscel, of human erythrocytes and of Staphylococcus aureus. A ratio of 1:5 was found to be necessary for the adenylate kinase from tobacco leaves and spinach chloroplasts, and a ratio of 2:1 was needed for suppressing the adenylate kinase from bovine liver mitochondria, human kidney homogenate and from Escherichia coli. Ap5A appears not to be metabolized in any of the above extracts. These results indicate that Ap5A can be used for evaluating the contribution of adenylate kinase to the production of ATP fro ADP in energy-transducing systems. 5. Contaminating adenylate kinase can be inhibited by a concentration of Ap5A which does not interfere in the study of many (phospho)kinases and ATPases. The applications of Ap5A in the assay for nucleoside diphosphokinase and in the study of mechanical and biochemical properties of contractile proteins are representative examples. The use of Ap5A makes it possible to study the effect of ADP per se in such systems. 6. Sepharose-bound Ap5A was used for removing traces of adenylate kinase from samples of myosin and creatine kinase. 7. In the presence of Ap5A the activity of creatine kinase was measured in hemolytic serum of venous blood, in plasma of capillary blood and in samples of whole blood after complete hemolysis had been induced. The clinical significance of these findings are shown for cases of myocardial infarction and muscular dystrophy.