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.
Published Version
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