Phosphorothioate analogues of 2',5'-oligoadenylate. Activation of 2',5'-oligoadenylate-dependent endoribonuclease by 2',5'-phosphorothioate cores and 5'-monophosphates.
Phosphorothioate analogues of 2',5'-oligoadenylate. Activation of 2',5'-oligoadenylate-dependent endoribonuclease by 2',5'-phosphorothioate cores and 5'-monophosphates.
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2,5-寡聚腺苷酸的硫代磷酸酯类似物。
DOI：
10.1021/bi00396a040
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发表时间：
1987
期刊：
影响因子：
2.9
通讯作者：
Pfleiderer,W
中科院分区：
文献类型：
--
作者：
Karikó,K;Li,SW;SobolJr,RW;Suhadolnik,RJ;Charubala,R;Pfleiderer,W
结果为了更清楚地了解 RNase L 水解 RNA 的结合和激活过程，对 2-5A 分子进行了立体化学修饰。利用这些2',5'-硫代磷酸酯的研究结果进一步阐明了2-5A分子中参与RNase L的结合和激活的区域。2'的5'-单磷酸化。具有 T4 多核苷酸激酶的 5'-硫代磷酸三聚体核心。 2/,5'-硫代磷酸酯三聚体核心的5'-单磷酸已经用ATP和T4多核苷酸激酶合成。通过 HPLC 分析确定 5'-单磷酸盐的形成（图 1）。使用化学和酶促磷酸化的 2', 5/-硫代磷酸酯三聚体 S'-单磷酸酯获得了相同的实验 HPLC 保留时间和电荷分离。随后用S'-核苷酸酶水解成核心。四个 2\5'-硫代磷酸酯三聚体核心的磷酸化取决于磷酸二酯主链的立体构型（表 I）。 5'-磷酸化对寡聚物更有效
ResultsTo understand more clearly the binding and activation processes by which RNase L hydrolyzes RNA, stereochemical modifications have been made in the 2-5A molecule. The results of studies utilizing these 2', 5'-phosphorothioates have further elucidated the regions of the 2-5 A molecule involved in binding to and activation of RNase L. 5'-Monophosphorylation of 2'. 5'-Phosphorothioate Trimer Cores with T4 Polynucleotide Ki tase. The 5'-monophosphates of the 2/, 5'-phosphorothioate trimer cores have been synthesized with ATP and T4 polynucleotide kinase. The formation of the 5'-monophosphates was determined by HPLC analyses (Figure 1). Identical experimental HPLC retention times and charge separations were obtained with the chemically and enzymatically phosphorylated 2', 5/-phosphorothioate trimer S'-monophosphates. Subsequent hydrolysis to cores was accomplished with S'-nucleotidase. The phosphorylation of the four 2\5'-phosphorothioate trimer cores was dependent on the stereoconfiguration in the phosphodiester backbone (Table I). 5'-Phosphorylation was more effective on oligomers