Zemlicka J
Biochemistry. 1980 Jan 8;19(1):163-8. doi: 10.1021/bi00542a025.
The chemical synthesis of the tital bridged trinucleoside diphosphates 3e and 3f along with the corresponding dinucleoside phosphates 3c and 3d is described. Bridged nucleosides 3a and 3b gave on treatment with triethyl orthoformate in the presence of p-toluenesulfonic acid in dimethylformamide the cyclic orthoesters 2a and 2b. Condensation of 2a and 2b with N,2',5'-O-triacetylcytidine 3'-phosphate (1) using dicyclohexylcarbodiimide in pyridine afforded after deblocking and chromatographic separation products 3c-f. The latter were readily degraded with pancreatic RNase, but 3c and 3e were completely resistant toward snake venom phosphodiesterase whereas 3d and 3f were digested to the extent of 65 and 43%, respectively. The major product of degradation of 3f with phosphodiesterase was compound 3d resulting from the combined action of phosphodiesterase and contaminating phosphomonoesterase. The results are explained in terms of stacking of terminal bridge nucleoside units in 3c-f. The implications of these findings for the function of snake venom phosphodiesterase are discussed.
本文描述了钛桥连三核苷二磷酸3e和3f以及相应的二核苷磷酸3c和3d的化学合成。桥连核苷3a和3b在对甲苯磺酸存在下于二甲基甲酰胺中用原甲酸三乙酯处理后得到环状原酸酯2a和2b。在吡啶中使用二环己基碳二亚胺使2a和2b与N,2',5'-O-三乙酰胞苷3'-磷酸(1)缩合,经脱保护和色谱分离后得到产物3c - f。后者易被胰核糖核酸酶降解,但3c和3e对蛇毒磷酸二酯酶完全耐受,而3d和3f分别被消化65%和43%。用磷酸二酯酶降解3f的主要产物是由磷酸二酯酶和污染的磷酸单酯酶共同作用产生的化合物3d。根据3c - f中末端桥连核苷单元的堆积情况对结果进行了解释。讨论了这些发现对蛇毒磷酸二酯酶功能的影响。