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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.

nucleic acids research

Procedure for C2 deuteration of nucleic acids and determination of Aχ31 psedouridine conformation by nuclear overhausere effect in yeast tRNA<sup>Phe</sup>

Procedure for C2 deuteration of nucleic acids and determination of Aχ31 psedouridine conformation by nuclear overhausere effect in yeast tRNAPhe

Procedure for C2 deuteration of nucleic acids and determination of Aχ31 psedouridine conformation by nuclear overhausere effect in yeast tRNA^Phe