Structures, ionization equilibria, and tautomerism of 6-oxopurines in solution.
6-Oxopurine and its analogues form an important class of biological molecules that include nucleobases and their precursors and are substrates of a wide range of enzymes. Solution structures of purines have been debated in the literature because of the many possible tautomers and protonation states in which they can exist in solution. Substitutions on the pyrimidine and imidazole rings alter tautomerization and protonation equilibria, and as a consequence, the solution compositions and structures of closely related analogues can be significantly different. We have obtained resonance Raman spectra of 6-oxopurines: hypoxanthine, xanthine, their riboside phosphates, guanine monophosphate in the protonated and deprotonated forms with UV excitation at 260 nm. The species present in solution under different pH conditions were identified by isotopic labeling with deuterium as well as by comparison with extensive density functional theoretical calculations. At physiological pH, while N7H and N9H tautomeric forms of hypoxanthine exist in equilibrium, in xanthine, the additional carbonyl group at C2 shifts the equilibrium in favor of the N7H tautomer. The corresponding nucleotide of xanthine, xanthosine monophosphate, on the other hand, is in the anionic form (pK(a) 5.5). We find that Raman spectra show systematic shifts with change in the protonation state and substitution on the ring. In general, deprotonation of the neutral molecule is marked by a downshift in the observed Raman wavenumbers, and protonation is accompanied by an upshift.
Gogia
Spriha
S
National Centre for Biological Sciences, GKVK Campus, Bellary Road, Bangalore 560065, India.
Jain
Ankur
A
Puranik
Mrinalini
M
D006863:Hydrogen-Ion Concentration; D007477:Ions; D007536:Isomerism; D015394:Molecular Structure; D011522:Protons; D011688:Purinones / Q000737:chemistry*; D012996:Solutions; D013059:Spectrum Analysis, Raman
D007477:Ions; D011522:Protons; D011688:Purinones; D012996:Solutions