Several properties of biologically-important furanoses (i.e., solution composition and conformation, and kinetics of ring-opening and ring-closing reactions (tautomerization) will be studied. The experimental approach involves: A. Synthesis of structural variants of furanoses and furanose phosphates in all ring configurations to permit structure-function comparisons. Compounds will be labeled with stable isotopes (13C, 2H) to facilitate examination by high-field NMR spectroscopy. B. Identification and quantitation of the tautomeric forms present in aqueous solutions of furanoses by NMR, with particular attention given to carbonyl and hydrate forms. C. Measurement and conformational interpretation of three-bond 1H-1H, 13C-1H and 13C-13C coupling constants and spin-lattice (T1) relaxation times (13C, 1H) in furanosyl rings. Relaxation studies will involve the use of DESERT and dynamic NOE NMR methods to obtain absolute 1H-1H internuclear distances, from which conformation will be assessed. Preferred ring conformations deduced by NMR will be compared to those predicted by MO calculations. D. Measurement and interpretation of unidirectional rate constants of furanose tautomerization by NMR. Activation parameters will be measured for each component reaction. Solution studies of simple furanoses will be extended to the ribonucleosides and oligoribonucleotides. Several GC-rich oligoribonucleotides, having base sequences identical to those found in the stems of E. coli tRNAglu, will be prepared containing perdeutero-ribofuranosyl residues to facilitate NMR studies of their solution structures. The proposed studies are a prerequisite to the pursuit of the longer-term objective of determining the molecular basis for the binding specificity between nucleic acids and proteins, and on elucidating the effect of sugar tautomerization on metabolic regulation.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM033791-03
Application #
3283830
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1984-12-01
Project End
1988-01-31
Budget Start
1986-12-01
Budget End
1988-01-31
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost
Name
University of Notre Dame
Department
Type
Schools of Arts and Sciences
DUNS #
824910376
City
Notre Dame
State
IN
Country
United States
Zip Code
46556
Wu, J; Serianni, A S (1994) 13C-labeled oligodeoxyribonucleotides: a solution study of a CCAAT-containing sequence at the nuclear factor I recognition site of human adenovirus. Biopolymers 34:1175-86
Podlasek, C A; Serianni, A S (1994) 19F and 13C NMR studies of polyol metabolism in freeze-tolerant pupae of Hyalophora cecropia. J Biol Chem 269:2521-8
Duker, J M; Serianni, A S (1993) (13C)-substituted sucrose: 13C-1H and 13C-13C spin coupling constants to assess furanose ring and glycosidic bond conformations in aqueous solution. Carbohydr Res 249:281-303
Wu, J; Serianni, A S; Bondo, P B (1992) Multiply 13C-substituted monosaccharides: synthesis of D-(1,5,6-13C3)glucose and D-(2,5,6-13C3)glucose. Carbohydr Res 226:261-9
Wu, J; Serianni, A S (1992) Isotope-edited 1D and 2D n.m.r. spectroscopy of 13C-substituted carbohydrates. Carbohydr Res 226:209-18
Wu, J A; Serianni, A S (1991) D-Penturonic acids: solution studies of stable-isotopically enriched compounds by 1H- and 13C-n.m.r. spectroscopy. Carbohydr Res 210:51-70
Snyder, J R; Serianni, A S (1991) Deoxygenated and alkylated furanoses: Thorpe-Ingold effects on tautomeric equilibria and rates of anomerization. Carbohydr Res 210:21-38
Vuorinen, T; Serianni, A S (1991) Synthesis of D-erythro-2-pentulose and D-threo-2-pentulose and analysis of the 13C- and 1H-n.m.r. spectra of the 1-13C- and 2-13C-substituted sugars. Carbohydr Res 209:13-31
Wu, J; Serianni, A S (1991) Ring-opening kinetics of the D-pentofuranuronic acids. Carbohydr Res 211:207-17
Vuorinen, T; Serianni, A S (1990) 13C-substituted pentos-2-uloses: synthesis and analysis by 1H- and 13C-n.m.r. spectroscopy. Carbohydr Res 207:185-210

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