The rate of lactate production from glucose in hearts is not altered by per-deuteration of glucose

Abstract

This study was designed to determine whether perdeuterated glucose experiences a kinetic isotope effect (KIE) as glucose passes through glycolysis and is further oxidized in the tricarboxylic acid (TCA) cycle. Metabolism of deuterated glucose was investigated in two groups of perfused rat hearts. The control group was supplied with a 1:1 mixture of [U-¹³C₆]glucose and [1,6-¹³C₂]glucose, while the experimental group received [U-¹³C₆,U-²H₇]glucose and [1,6-¹³C₂]glucose. Tissue extracts were analyzed by ¹H, ²H and proton-decoupled ¹³C NMR spectroscopy. Extensive ²H-¹³C scalar coupling plus chemical shift isotope effects were observed in the proton-decoupled ¹³C NMR spectra of lactate, alanine and glutamate. A small but measureable (∼8%) difference in the rate of conversion of [U-¹³C₆]glucose vs. [1,6-¹³C₂]glucose to lactate, likely reflecting rates of CC bond breakage in the aldolase reaction, but conversion of [U-¹³C₆]glucose versus [U-¹³C₆,U-²H₇]glucose to lactate did not differ. This shows that the presence of deuterium in glucose does not alter glycolytic flux. However, there were two distinct effects of deuteration on metabolism of glucose to alanine and oxidation of glucose in the TCA. First, alanine undergoes extensive exchange of methyl deuterons with solvent protons in the alanine amino transferase reaction. Second, there is a substantial kinetic isotope effect in metabolism of [U-¹³C₆,U-²H₇]glucose to alanine and glutamate. In the presence of [U-¹³C₆,U-²H₇]glucose, alanine and lactate are not in rapid exchange with the same pool of pyruvate. These studies indicate that the appearance of hyperpolarized ¹³C-lactate from hyperpolarized [U-¹³C₆,U-²H₇]glucose is not substantially influenced by a deuterium kinetic isotope effect.

Keywords: Deuterium; Glucose metabolism; Heart; Kinetic isotope effect; NMR; Perfusion.