Morozov I A, Gambarian A S, Venkstern T V
Mol Biol (Mosk). 1984 Sep-Oct;18(5):1363-8.
tRNA(adenine-1-)-methyltransferase (EC 2.1.1.36) was isolated from the extreme thermophile Thermus thermophilus strain HB8. The specific activity of the enzyme is about 50 000 and the yield of activity more than 20%. The method of isolation consists of five steps and is valid for isolation of mg quantities of the enzyme. The purified protein preparation is practically homogeneous in SDS-gel electrophoresis, the position of the protein band corresponds to a molecular weight of 25 000. By gel filtration on Sephadex G-100 the molecular weight of the native protein was found to be 70 000. These data allow to suggest a subunit structure of the enzyme. The enzyme is highly thermostable and is most active at 80 degrees C. The only activity of the enzyme is to methylate A58 in the T psi X loop of tRNA.
从嗜热栖热菌HB8菌株中分离出了tRNA(腺嘌呤-1-)-甲基转移酶(EC 2.1.1.36)。该酶的比活性约为50000,活性回收率超过20%。分离方法包括五个步骤,适用于毫克量酶的分离。纯化后的蛋白质制剂在SDS-凝胶电泳中几乎是均一的,蛋白条带位置对应的分子量为25000。通过Sephadex G-100凝胶过滤发现天然蛋白的分子量为70000。这些数据表明了该酶的亚基结构。该酶具有高度的热稳定性,在80℃时活性最高。该酶的唯一活性是使tRNA的TψX环中的A58甲基化。