supK tRNA甲基化酶的tRNA底物鉴定
The identification of the tRNA substrates for the supK tRNA methylase.
作者信息
Pope W T, Brown A, Reeves R H
出版信息
Nucleic Acids Res. 1978 Mar;5(3):1041-57. doi: 10.1093/nar/5.3.1041.
Abstract
Purified preparations of the tRNA methylase deficient in supK strains of Salmonella typhimurium transfer methyl groups from S-adenosylmethionine (SAM) to at least two tRNA species, an alanine tRNA and a serine tRNA. The identity of the tRNA substrates for this enzyme was determined by a change in the elution position of the methyl-labeled tRNA from BND-cellulose columns before and after aminoacylation with a specific amino acid followed by derivatization of the free primary amino group with phenoxy- or naphthoxyacetate. The radioactive methyl group enzymatically added to these tRNAs is both acid and base labile and can be hydrolyzed to a volatile product at pHs above 7.5 and also at pH 1. The methylated 3'-nucleotide isolated from digested tRNA is a pyrimidine derivative and chromatographs like a modified uridylic acid. Its identity has not been established, but it is likely that it corresponds to the methyl ester of V, uridin-5-oxyacetic acid.
摘要
鼠伤寒沙门氏菌supK菌株中缺乏的tRNA甲基化酶的纯化制剂可将甲基从S-腺苷甲硫氨酸（SAM）转移至至少两种tRNA，即丙氨酸tRNA和丝氨酸tRNA。该酶的tRNA底物的身份是通过用特定氨基酸进行氨酰化，然后用苯氧基或萘氧基乙酸对游离伯氨基进行衍生化前后，甲基标记的tRNA从BND-纤维素柱上洗脱位置的变化来确定的。酶促添加到这些tRNA上的放射性甲基对酸和碱都不稳定，在pH高于7.5以及pH 1时均可水解为挥发性产物。从消化后的tRNA中分离出的甲基化3'-核苷酸是一种嘧啶衍生物，其色谱行为类似于修饰的尿苷酸。其身份尚未确定，但很可能它对应于V，尿苷-5-氧乙酸的甲酯。
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