酵母苯丙氨酸转运核糖核酸中核酸C2位氘代及通过核Overhauser效应测定ψ31假尿苷构象的方法
Procedure for C2 deuteration of nucleic acids and determination of A psi 31 pseudouridine conformation by nuclear Overhauser effect in yeast tRNAPhe.
作者信息
Roy S, Papastavros M Z, Redfield A G
出版信息
Nucleic Acids Res. 1982 Dec 20;10(24):8341-9. doi: 10.1093/nar/10.24.8341.
Abstract
Nuclear Overhauser effect (NOE) combined with semispecific deuteration provides a general strategy for identification of exchangeable protons in nucleic base pairs, and has been extended to NOEs involving purine C2 protons in tRNA. Deuterated tri-ethyl orthoformate was condensed with 5(4)-amino imidazole 4(5)-carboxamide to yield C2 deuterated hypoxanthine. C2 deuterated hypoxanthine was fed to a purine requiring mutant of yeast and C2 deuterated yeast tRNAPhe was isolated. This C2 deuterated tRNAPhe was used to identify A psi 31 and U8-A14. A psi 31 was found to be bonded through N1H. The utility of C2 deuteration in nucleic acid NMR is thus demonstrated.
摘要
核Overhauser效应（NOE）与半特异性氘代相结合，为鉴定核酸碱基对中可交换质子提供了一种通用策略，并且已扩展到涉及tRNA中嘌呤C2质子的NOE。用氘代原甲酸三乙酯与5(4)-氨基咪唑4(5)-甲酰胺缩合，得到C2氘代次黄嘌呤。将C2氘代次黄嘌呤喂给需要嘌呤的酵母突变体，分离得到C2氘代酵母苯丙氨酸tRNA。这种C2氘代苯丙氨酸tRNA用于鉴定Aψ31和U8-A14。发现Aψ31通过N1H键合。由此证明了C2氘代在核酸核磁共振中的实用性。
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