Glucosamine Research Today is a free monthly online journal that collates and summarizes the latest research about Glucosamine, including details on chondroitin sulfate, uses, effects, benefits, arthritis.

Monitoring enzyme-catalyzed production of glucosamine-6P by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: a new enzymatic assay for glucosamine-6P synthase.

Maillard LT, Guérineau V, Badet-Denisot MA, Badet B, Laprévote O, Durand P

Institut de Chimie des Substances Naturelles, UPR 2301‐CNRS, Avenue de la Terrasse, 91198 Gif‐sur‐Yvette, France.

A matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) method for quantification of D-glucosamine-6P (GlcN-6P) that allows the kinetic study of glucosamine-6P synthase (Glms) is presented. The present report describes the optimization of the different steps of a new enzymatic assay for Glms based on in situ N-acetylation of GlcN-6P and MALDI-TOFMS analysis using N-((13)C(2))acetylglucosamine-6P as internal standard. Since no isotopically substituted GlcN-6P was available, the N-((13)C(2))acetyl derivative, easily obtained from ((13)C(4))-acetic anhydride, was used as internal standard. Validation of the assay was achieved by measuring the fructose-6P Michaelis constant, in full agreement with reported values, and by studying the inhibition properties of arabinose-5P oxime. Copyright (c) 2006 John Wiley & Sons, Ltd.

Published 30 January 2006 in Rapid Commun Mass Spectrom, 20(4): 666-672.
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