DFT analysis of nitrogen isotopic reduction partition function ratios in proton exchange equilibria of glutamic acid species in water and its application to the estimation of nitrogen isotope fractionation.

Abstract

L-Glutamic acid (Glu) plays a pivotal role in amino acid metabolism in living organisms. Theoretical knowledge of how metabolism propagates the nitrogen isotope composition (δ15N) is essential for elucidating the mechanism of amino acid metabolism in vivo. In this paper, to estimate the nitrogen isotope fractionation involving Glu and its protonated/deprotonated species, their nitrogen isotopic reduction partition function ratios (RPFRs) were calculated at the apfd/6-311+G(2d,p) level of theory by the SCRF method. The results show that the RPFR values of the eight possible Glu species at 25 °C are: cation > zwitterion with α-COOH and γ-COO- > zwitterion with α-COO- and γ-COOH > anion with -NH3+ ≫ uncharged molecule > anion with -NH2 and γ-COO- > anion with -NH2 and α-COO- > di-anion. Several correlations between RPFR and bond distances were found. Most importantly, it was found that the shorter the distance of the (C-)N⋯H(-O) hydrogen bond (HB), the greater the RPFR value for that species. This correlation indicates that the tri-coordinate nitrogen atom of the amino group may take on the character of the tetra-coordinate nitrogen atom to some extent by forming a HB of this type. The nitrogen isotope exchange equilibrium between Glu and glycine/serine was evaluated at a typical physiological pH of 7.4. Calculations suggest that the equilibrium constant for the former is an increasing function of temperature from 0 to 100 °C, while the latter is a decreasing function of temperature over the same temperature range.