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- Title
Unambiguous assignment of the H3 S and H3 R deuterations of cerebral (2-<sup>13</sup>C) glutamate by <sup>13</sup>C NMR at 18.8 tesla.
- Authors
Rodrigues, Tiago B.; Violante, Inês R.; Cerdán, Sebastián
- Abstract
We used high-field 13C NMR (18.8 T) to assign unambiguously the isotopic shifts induced by the deuterium substitutions of the H3 proR and H3 proS hydrogens of (2-13C) glutamate in extracts of the brain from deuterated animals. Monodeuterated H3 R or H3 S glutamate diastereoisomers were produced stereospecifically either by chemical synthesis or by coupling the reactions of isocitrate dehydrogenase and aspartate aminotransferase in deuterated medium, respectively. We show that the (3 S-2H) or (3 R-2H) deuterations induce characteristic small (Δ2 = −0.058 parts per million (ppm)) or large (Δ2 = −0.071 ppm) vicinal isotopic shifts upfield of the perprotonated (2-13C) glutamate resonance (at 55.5 ppm). Isotopically shifted (2-13C, 3 S-2H) or (2-13C, 3 R-2H) glutamate singlets are conveniently observed by high-field 13C NMR in brain extracts from deuterated rats. Since the (3 S-2H) or (3 R-2H) glutamate diastereoisomers are produced stereospecifically by the cytosolic or mitochondrial isoforms of aconitase and isocitrate dehydrogenase, our results will facilitate the 13C NMR investigation of these enzymatic activities and their role in subcellular glutamate trafficking. Magn Reson Med 63:1088-1091, 2010. © 2010 Wiley-Liss, Inc.
- Publication
Magnetic Resonance in Medicine, 2010, Vol 63, Issue 4, p1088
- ISSN
0740-3194
- Publication type
Article
- DOI
10.1002/mrm.22277