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首页> 外文期刊>Biochemical Pharmacology >Synthesis of ATP derivatives of compounds of the mevalonate pathway (isopentenyl di- and triphosphate; geranyl di- and triphosphate, farnesyl di- and triphosphate, and dimethylallyl diphosphate) catalyzed by T4 RNA ligase, T4 DNA ligase and other ligases Potential relationship with the effect of bisphosphonates on osteoclasts.
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Synthesis of ATP derivatives of compounds of the mevalonate pathway (isopentenyl di- and triphosphate; geranyl di- and triphosphate, farnesyl di- and triphosphate, and dimethylallyl diphosphate) catalyzed by T4 RNA ligase, T4 DNA ligase and other ligases Potential relationship with the effect of bisphosphonates on osteoclasts.

机译：T4 RNA连接酶，T4 DNA连接酶和其他连接酶催化的甲羟戊酸途径化合物（异戊烯基二磷酸和三磷酸;香叶基二磷酸和三磷酸，法呢基二磷酸和三磷酸以及二甲基烯丙基二磷酸）的ATP衍生物的合成双膦酸盐对破骨细胞的作用。

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Compounds of the mevalonate pathway containing a terminal di- or triphosphate (mev-PP or mev-PPP) were tested as substrates of several enzyme ligases (T4 RNA ligase, T4 DNA ligase, firefly luciferase and other ligases) for the synthesis of ATP derivatives of the mev-pppA or mev-ppppA type. T4 RNA ligase, in the presence of ATP and the substrates: geranyl, farnesyl or isopentenyl triphosphates, and geranyl, farnesyl, dimethylallyl or isopentenyl diphosphates, all at 0.3 mM concentration, catalyzed the synthesis of the corresponding ATP derivatives at a relative rate of activity of: 7.6+/-1.4 mU/mg or 100%; 39%; 42%; 24%; 18%; 12% and 6%, respectively. Inhibition (%) of the synthesis by excess of substrate (0.8 mM vs. 0.3 mM) was observed with farnesyl diphosphate (99%); farnesyl triphosphate (96%) and geranyl triphosphate (32%). V(max), K(m), K(cat) and K(cat)/K(m) values were also determined. The K(cat)/K(m) values calculated were for: farnesyl triphosphate, 166; geranyl triphosphate, 52.2; farnesyl diphosphate, 12.1; geranyl diphosphate, 8.6; isopentenyl triphosphate, 6.7; dimethylallyl diphosphate, 3.1 and isopentenyl diphosphate, 0.9. Similar results were obtained with T4 DNA ligase. The above-mentioned compounds were also substrates of firefly luciferase synthesizing the mev-pppA or mev-ppppA derivatives. In our hands, neither the acyl- or acetyl-CoA synthetases nor the ubiquiting activating enzyme (E1) catalyzed the synthesis of ATP derivatives of these compounds. The results here presented could be related with the mechanism of action of bisphosphonates on osteoclasts or tumor cells.
机译：含有末端二磷酸或三磷酸的甲羟戊酸途径的化合物（mev-PP或mev-PPP）已作为几种用于合成ATP衍生物的酶连接酶（T4 RNA连接酶，T4 DNA连接酶，萤火虫荧光素酶和其他连接酶）的底物进行了测试mev-pppA或mev-ppppA类型的。 T4 RNA连接酶在ATP和底物的存在下：浓度为0.3 mM的香叶基，法呢基或异戊烯基三磷酸酯和香叶基，法呢基，二甲基烯丙基或异戊烯基二磷酸酯均以相对活性速率催化了相应ATP衍生物的合成of：7.6 +/- 1.4 mU / mg或100％; 39％; 42％; 24％; 18％;分别为12％和6％。用法呢基二磷酸酯（99％）观察到过量底物（0.8 mM vs. 0.3 mM）对合成的抑制（％）;法呢基三磷酸酯（96％）和香叶基三磷酸酯（32％）。还确定了V（max），K（m），K（cat）和K（cat）/ K（m）值。计算的K（cat）/ K（m）值是：三磷酸法呢酯166;三磷酸香叶基酯52.2;法呢基二磷酸酯，12.1;香叶基二磷酸8.6;异戊烯基三磷酸6.7;二磷酸二甲基烯丙酯3.1和0.9异戊烯二磷酸。 T4 DNA连接酶获得了相似的结果。上述化合物也是萤火虫荧光素酶的底物，其合成了mev-pppA或mev-ppppA衍生物。在我们手中，酰基-或乙酰基-CoA合成酶或遍在的活化酶（E1）都不催化这些化合物的ATP衍生物的合成。这里介绍的结果可能与双膦酸盐对破骨细胞或肿瘤细胞的作用机制有关。

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