par Malaisse Lagae, Francine ;Zahner, Dagmar ;Sener, Abdullah ;Malaisse, Willy
Référence Bulletin des Sociétés chimiques belges, 101, 2, page (159-164)
Publication Publié, 1992
Référence Bulletin des Sociétés chimiques belges, 101, 2, page (159-164)
Publication Publié, 1992
Article révisé par les pairs
Résumé : | An enzymic method is presented for the preparation of 14C‐labelled and C1‐deuterated D‐glucose, to be used as a radioactive tracer in metabolic studies dealing with the fate of D‐[1‐2H]glucose. The procedure takes advantage of the deuteration of 14C‐labelled D‐fructose 6‐phosphate during exposure to phosphomannoisomerase in D2O. It further includes the conversion of the deuterated hexose monophosphates to D‐fructose 1,6‐bisphosphate, its isolation and hydrolysis to D‐fructose 6‐phosphate and the partial conversion of the latter ester to D‐glucose 6‐phosphate, which is then isolated by ion exchange chromatography and hydrolyzed to unesterified D‐glucose. The extent of deuteration on the C1 of D‐glucose was judged, after conversion to D‐glucose 6‐phosphate, from the isotopic discrimination in reaction velocity exerted towards the C1‐deuterated ester by glucose‐6‐phosphate dehydrogenase. The degree of isotopic discrimination was virtually identical to that found with D‐glucose 6‐phosphate prepared from pure D‐[1‐2H]glucose. Copyright © 1992 Wiley‐VCH Verlag GmbH & Co. KGaA, Weinheim |