T4 RNA LIGASE MEDIATED PREPARATION OF NOVEL CHEMICALLY MISACYLATED TRANSFER-RNA PHES
T4 RNA LIGASE MEDIATED PREPARATION OF NOVEL CHEMICALLY MISACYLATED TRANSFER-RNA PHES
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DOI：
10.1021/bi00302a020
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发表时间：
1984-01-01
期刊：
影响因子：
2.9
通讯作者：
HECHT, SM
中科院分区：
文献类型：
--
作者：
HECKLER, TG;CHANG, LH;HECHT, SM
噬菌体T4 RNA连接酶用于大肠杆菌tRNAPHe缺失胞苷-75和腺苷-76 （tRNAPHe- coh；受体低聚物）与pCpA的几种化学酰化衍生物（供体低聚物）的缩合。在deae -纤维素和苯甲酰化的deae -纤维素上进行色谱分析，得到了20-65%的化学misacylated tRNAPHe。对化学错酰化tRNA的表征是通过大肠杆菌苯丙酰-tRNA合成酶在对错误的氨基酸进行化学去酰化后，将化学错酰化的tRNAPHe与苯丙氨酸酶解氨基酰化，比较Cu2+溶液对化学和酶解制备的n -乙酰基- l-苯丙酰-tRNAPHe的水解效果，以及测量化学错酰化产生的tRNA物种的色谱行为。
Phage T4 RNA ligase was employed for the condensation of Escherichia coli tRNAPHe missing cytidine-75 and adenosine-76 (tRNAPHe-COH; the acceptor oligomer) with each of several chemically acylated derivatives of pCpA (the donor oligomer). The resulting chemically misacylated tRNAPHe were obtained in 20-65% yields following chromatographic workup on DEAE-cellulose and benzoylated DEAE-cellulose. Characterization of the chemically misacylated tRNA was accomplished by enzymatic reaminoacylation of chemically misacylated tRNAPHe with phenylalanine by E. coli phenylalanyl-tRNA synthetase following chemical deacylation of the incorrect amino acid, comparison of the hydrolytic effects of Cu2+ solutions on chemically and enzymatically prepared samples of N-acetyl-L-phenylalanyl-tRNAPHe, and measurement of the chromatographic behavior of the tRNA species derived from chemical misacylation.