Data on the phosphorylation state of the catalytic serine of enzymes in the α-D-phosphohexomutase superfamily

Abstract

Most enzymes in the α-D-phosphohexomutase superfamily catalyze the reversible conversion of 1- to 6-phosphosugars. They play important roles in carbohydrate and sugar nucleotide metabolism, and participate in the biosynthesis of polysaccharides, glycolipids, and other exoproducts. Mutations in genes encoding these enzymes are associated with inherited metabolic diseases in humans, including glycogen storage disease and congenital disorders of glycosylation. Enzymes in the superfamily share a highly conserved active site serine that participates in the multi-step phosphoryl transfer reaction. Here we provide data on the effects of various phosphosugar ligands on the phosphorylation of this serine, as monitored by electrospray ionization mass spectrometry (ESI-MS) data on the intact proteins. We also show data on the longevity of the phospho-enzyme under various solution conditions in one member of the superfamily from Pseudomonas aeruginosa, and present inhibition data for several ligands. These data should be useful for the production of homogeneous samples of phosphorylated or unphosphorylated proteins, which are essential for biophysical characterization of these enzymes.

Genetic validation of Aspergillus fumigatus phosphoglucomutase as a viable therapeutic target in invasive aspergillosis

“…S1 A ). The M w difference (82 Da) between the two protein species likely corresponds to the M w of a phosphoryl group (79 Da), which is compatible with the observation that intracellular ATP/Glc-1,6-2P phosphorylates a catalytic serine in the active site of phosphohexomutases ( 78 , 79 , 85 ). The enzymatic activity of Af PGM for Glc-1P was determined using a Glc-6P dehydrogenase (G6PDH) coupled enzyme assay.…”

Asp263 missense variants perturb the active site of human phosphoglucomutase 1

“…To test this biochemically, hydrolysis of P‐Ser117 over time was assessed for WT PGM1 and both the D263Y and D263G variants. (Both time and increased temperature are associated with a reduction in the level of P‐Ser of related enzymes ). Fully phosphorylated (> 90%) versions of WT PGM1 and the D263Y/G variants were prepared using established protocols (Experimental procedures).…”

Phosphorylation-Dependent Effects on the Structural Flexibility of Phosphoglucosamine Mutase from Bacillus anthracis

“…The recombinant BaPNGM purifies with Ser100 in its dephosphorylated state. , Samples phosphorylated at Ser100 were prepared as in Experimental Procedures. Phosphorylation status was verified by electrospray ionization mass spectrometry (ESI-MS) (Figure S1) prior to use.…”