Determination of the anomeric configuration of glycosyl esters of nucleoside pyrophosphates and polyisoprenyl phosphates by fast-atom bombardment tandem mass spectrometry.

Collision-induced dissociation of the deprotonated molecules of glycosyl esters of nucleoside pyrophosphates and polyisoprenyl (dolichyl and polyprenyl) phosphates results in distinct fragmentation patterns that depend on cis-trans configuration of the phosphodiester and 2″ (or 2', respectively)-hydroxyl groups of the glycosyl residue. At the collision-offset voltage of 0. 5 V, sugar nucleotides with cis configuration produce only one very abundant fragment of nucleoside monophosphate, whereas compounds with trans configuration give weak signals for nucleoside di- and mono-phosphates and their dehydration products. These fragmentation patterns are largely preserved at higher collision energy, with the exception that, for sugar nucleotides with trans configuration, the characteristic signals are much more abundant and a novel diagnostic fragment of [ribosyl(deoxyribosyl)-5'-P2O5 - H](-) is generated. In the case of polyisoprenyl-P-sugars, polyisoprenyl phosphate ion is the only fragment observed for compounds with trans configuration, whereas in compounds with cis configuration, this ion is accompanied by another abundant fragment, which is derived from the cleavage across the sugar ring and corresponds to [polyisoprenyl-PO4-(C2H3O)](-). The relative intensity ratio of the latter ion to the [polyisoprenyl-HPO4](-) ion is close to 1 for compounds with cis configuration, but it is only about 0. 01 for compounds with trans configuration. This ratio may serve, therefore, as a diagnostic value for determination of the anomeric configuration of glycosyl esters of polyisoprenyl phosphates. It is proposed that the observed differences in fragmentation patterns of cis-trans sugar nucleotides and polyisoprenyl-P-sugars could be explained in terms of kinetic stereoelectronic effect, and a speculative mechanism of fragmentation of compounds with trans configuration is presented. For compounds with cis configuration, formation of a hydrogen bond between the C-2″(2') hydroxyl and the phosphate group could play a crucial role in directing the specific fragmentation reactions. Consequently, the described empirical rules would hold only for compounds that have a free 2″(2')-hydroxyl group and no alternative charge location. Owing to its simplicity, sensitivity, and tolerance of impurities, fast-atom bombardment-tandem mass spectrometry represents a suitable method for determination of the anomeric linkage of glycosyl esters of nucleoside pyrophosphates and polyisoprenyl phosphates if the absolute configuration of glycosyl residue is known and the compound fulfills the above-mentioned requirements.