Nedospasov A A, Khomutov R M, Gambarian A S, Morozov I A, Venkstern T V
Mol Biol (Mosk). 1978 Sep-Oct;12(5):1104-11.
A new type of sorbents for affinity chromatography is suggested and used to purify tRNA methylases. tRNA was immobilized on aminooxybutylcellulose via the oxidized 3'-end. In order to bind other enzymes specific for nucleic acids in general, e. g. nucleases, and to achieve a higher degree of purification the crude enzyme preparation was treated with rRNA immobilized on aminooxybutycellulose. The sequential application of two sorbents mentioned allows to get an approximately two hundred fold purification of total tRNA methylases. In a separate experiment the possibility of individual tRNA methylase fractionation by means of elution with a NACl gradient was shown; the degree of purification for some methylases was more than a thousand fold.
提出了一种新型的亲和色谱吸附剂,并用于纯化tRNA甲基化酶。tRNA通过氧化的3'-末端固定在氨氧基丁基纤维素上。为了结合一般对核酸具有特异性的其他酶,例如核酸酶,并实现更高程度的纯化,粗酶制剂用固定在氨氧基丁基纤维素上的rRNA处理。依次应用上述两种吸附剂可使总tRNA甲基化酶得到约200倍的纯化。在一个单独的实验中,展示了通过用NaCl梯度洗脱来分离单个tRNA甲基化酶的可能性;某些甲基化酶的纯化程度超过了1000倍。