Wierzbicka H, Jakubowski H
Nucleic Acids Res. 1975 Jan;2(1):101-11. doi: 10.1093/nar/2.1.101.
tRNA methyltransferases from extract of yellow lupin seeds were purified over 300-fold by the methods based on hydrophobic and affinity chromatography. However, in the most active fractions the methylating enzymes were over 2000 purified. The purified enzyme fractions catalysed the formation of 1-methyladenine and 5-methylcytosine using E. coli B and B. subtilis tRNAs as substrates and S-adenosylmethionine as the methyl donor. They were unable to methylate their own endogenous tRNA but they were capable of methylating tRNA of some other lupinus species. Whereas the patterns of methylated constituents of tRNA of some other lupinus and B. subtilis were quite similar, they differed considerably from those obtained with lupin species tRNAs. Some properties of purified methyltransferases from yellow lupin seeds have been described.
通过基于疏水色谱和亲和色谱的方法,从黄羽扇豆种子提取物中纯化出的tRNA甲基转移酶的纯化倍数超过300倍。然而,在活性最高的组分中,甲基化酶的纯化倍数超过了2000倍。纯化后的酶组分以大肠杆菌B和枯草芽孢杆菌的tRNA为底物、S-腺苷甲硫氨酸为甲基供体,催化形成1-甲基腺嘌呤和5-甲基胞嘧啶。它们无法甲基化自身的内源性tRNA,但能够甲基化其他一些羽扇豆属物种的tRNA。虽然其他一些羽扇豆属和枯草芽孢杆菌的tRNA甲基化成分模式相当相似,但与羽扇豆属物种tRNA的甲基化成分模式有很大差异。本文描述了从黄羽扇豆种子中纯化出的甲基转移酶的一些特性。