{Reference Type}: Journal Article
{Title}: Measuring Phosphoglycolate Phosphatase Activity Using NMR Detection of Glycolate.
{Author}: Mauve C;Oury C;Glab N;Hodges M;
{Journal}: Methods Mol Biol
{Volume}: 2792
{Issue}: 0
{Year}: 2024
暂无{DOI}: 10.1007/978-1-0716-3802-6_2
{Abstract}: Besides the historical and traditional use of nuclear magnetic resonance (NMR) spectroscopy as a structure elucidation tool for proteins and metabolites, its quantification ability allows the determination of metabolite amounts and therefore enzymatic activity measurements. For this purpose, 1H-NMR with adapted water pulse pre-saturation sequences and calibration curves with commercial standard solutions can be used to quantify the photorespiratory cycle intermediates, 2-phosphoglycolate and glycolate, associated with the phosphoglycolate phosphatase reaction. The intensity of the 1H-NMR signal of glycolate produced by the activity of purified recombinant Arabidopsis thaliana PGLP1 can therefore be used to determine PGLP1 enzymatic activities and kinetic parameters.