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Nucleic Acids Research, 1982, Vol. 10, No. 24 8341-8349
© 1982

CHEMISTRY

Procedure for C2 deuteration of nucleic acids and determination of A{chi}31 psedouridine conformation by nuclear overhausere effect in yeast tRNAPhe

Siddhartha Roy*, Mary Z. Papastavros* and Alfred G. Redfield+

+Department of Biochemistry, and physics, and Rosenstiel Basic Medical Sciences Research Center, Brandeis University Waltham, MA02254, USA *DepartmentofBiochemistry

Received July 1, 1982. Accepted October 18, 1982.

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)-aminoimidazole 4(5)-carboxamide to yield C2 deuterated hypoxanthine. C2 deuterated hypoxanthine was fed to a purine requiring mutant of yeast and C2 deuterated tRNAPhe was isolated. This C2 deuterated tRNAPhe was used to identify A{chi}31 and U8-A14. A{chi}31 was found to be bonded through NlH. The utility of C2 deuteration in nucleic acid NMR is thus demonstrated.


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