Salomon R, Giveon D, Kimhi Y, Littauer U Z
Biochemistry. 1976 Nov 30;15(24):5258-62. doi: 10.1021/bi00669a010.
Affinity chromatography on anti-Y (Y is a tricyclic imidazopurine to which is attached a complex four-carbon side chain) antibody immobilized to Sepharose was used to determine the proportion of rat liver tRNAPhe species containing the peroxy Y-nucleoside. Unfractionated Unfractionated mammalian tRNA was aminoacylated with labeled phenylalanine. The phenylalanyl-tRNA was then chemically acetylated to yield N-acetylphenylalanyl-tRNA. When this preparation was applied to the antibody column, between 6-10% of the radioactivity was not bound to the column, indicating a deficiency of peroxy Y-nuceloside in a minor isoaccepting tRNAPhe species. In contrast to normal tissues (including embryonic tissue), about 85% of the tRNAPhe from mouse neuroblastoma C-1300 or N-18 tumors lack the peroxy Y-base, a property which is not affected by tumor age. Rat liver labeled N-acetylphenylalanyl-tRNA preparations were resolved on Plaskon chromatography (RPC-5) into two minor peaks closely followed by a mojor component. A high proportion of the two minor tRNAPhe species was unable to bind to anti-Y antibodies. Upon mild acid treatment, the minor and major tRNAPhe species eluted simultaneously from Plaskon columns, at a much reduced salt concentration. These results would indicate that the two minor tRNAPhe species from rat liver as well as the major component contain a tricyclic imidazopurine base that differs from each other in its side chain. About 85% of the N-acetylphenylalanyl-tRNA from neuroblastoma was resolved by Plaskon chromatography as an early eluting peak. The position of this major neuroblastoma tRNAPhe species was not altered by mild acid treatment, and its elution position from the column almost coincides with that of acid-treated normal rat liver tRNAPhe. The latter results would suggest that most of the tRNAPhe chains from neuroblastoma lack the tricyclic imidazopurine of normal rat liver tRNAPhe, but are very close if not identical in primary nucleotide sequence.
将抗Y（Y是一种三环咪唑嘌呤，连接有复杂的四碳侧链）抗体固定在琼脂糖珠上，通过亲和层析法来测定大鼠肝脏中含有过氧Y-核苷的苯丙氨酸tRNA种类的比例。未分级分离的哺乳动物tRNA用标记的苯丙氨酸进行氨酰化。然后将苯丙氨酰-tRNA进行化学乙酰化，得到N-乙酰苯丙氨酰-tRNA。当将该制剂应用于抗体柱时，6%-10%的放射性未与柱结合，表明在一种次要的同功受体苯丙氨酸tRNA种类中缺乏过氧Y-核苷。与正常组织（包括胚胎组织）相比，来自小鼠神经母细胞瘤C-1300或N-18肿瘤的约85%的苯丙氨酸tRNA缺乏过氧Y碱基，这一特性不受肿瘤年龄的影响。大鼠肝脏标记的N-乙酰苯丙氨酰-tRNA制剂在普拉斯康色谱法（RPC-5）上分离为两个小峰，紧随其后的是一个主要成分。这两种次要的苯丙氨酸tRNA种类中有很大比例不能与抗Y抗体结合。经过温和酸处理后，次要和主要的苯丙氨酸tRNA种类在盐浓度大幅降低的情况下同时从普拉斯康柱上洗脱下来。这些结果表明，来自大鼠肝脏的两种次要苯丙氨酸tRNA种类以及主要成分都含有一种三环咪唑嘌呤碱基，它们的侧链彼此不同。约85%的来自神经母细胞瘤的N-乙酰苯丙氨酰-tRNA在普拉斯康色谱法上作为一个早期洗脱峰被分离出来。这种主要的神经母细胞瘤苯丙氨酸tRNA种类的位置在温和酸处理后没有改变，其从柱上的洗脱位置几乎与酸处理后的正常大鼠肝脏苯丙氨酸tRNA的洗脱位置一致。后一个结果表明，神经母细胞瘤的大多数苯丙氨酸tRNA链缺乏正常大鼠肝脏苯丙氨酸tRNA的三环咪唑嘌呤，但在一级核苷酸序列上即使不完全相同也非常接近。
