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High yield synthesis of 14 C labelled intermediates of the L‐type pentose pathway: Octulose mono‐ and bisphosphates, sedoheptulose 1,7‐bisphosphate and D‐arabinose 5‐phosphate
Author(s) -
Arora Krishan K.,
Macleod John K.,
Williams John F.
Publication year - 1987
Publication title -
journal of labelled compounds and radiopharmaceuticals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.432
H-Index - 47
eISSN - 1099-1344
pISSN - 0362-4803
DOI - 10.1002/jlcr.2580240210
Subject(s) - chemistry , sugar phosphates , pentose , transketolase , pentose phosphate pathway , arabinose , dihydroxyacetone phosphate , aldolase a , transaldolase , stereochemistry , biochemistry , enzyme , xylose , fermentation , glycolysis
Methods for the enzymic synthesis, isolation, purification and analysis of the 14 C labelled intermediates that are characteristic of the L‐type pentose phosphate pathway are described. These are D‐ glycero D‐ ido octulose 1, 8‐bisphosphate and 8‐monophosphate; D‐ glycero D‐ altro octulose mono‐ and bisphosphates; sedoheptulose 1,7‐bisphosphate and D‐arabinose 5‐phosphate. The procedures for the preparation of octulose‐ and sedoheptulose bisphosphates are based on the aldolase‐catalyzed condensation of dihydroxyacetone phosphate with appropriately 14 C‐labelled aldo pentose 5‐phosphates and erythrose 4‐phosphate respectively. D‐ Glycero D‐ ido octulose 8‐phosphate and its altro epimer were prepared in transketolase catalyzed reactions involving β‐hydroxypyruvate and appropriately 14 C‐labelled aldohexose 6‐phosphates. [U‐ 14 C] D‐arabinose 5‐phosphate was synthesized by reacting [U‐ 14 C] glucosamine 6‐phosphate with ninhydrin. The authenticity of each of the 14 C‐labelled sugar phosphates was confirmed using a variety of chromatographic methods, enzymatic analytic methods and NMR spectroscopy. The 14 C labelled compounds are used in investigations involving the search for the identity of the pentose pathway in tissues in vitro , for the measurement of L‐type pathway enzyme reactions and for testing the mechanistic predictions of the L‐type pathway in vitro .