Glucose-6-phosphate dehydrogenase (G6PDH) can initiate the glucose-6-phosphate (G6P) shunt around the Calvin-Benson cycle. To understand the regulation of flux through this pathway， we have characterized the biochemical parameters and redox regulation of the three functional plastidic isoforms of G6PDH. When purified， recombinant proteins were measured， all three exhibited significant substrate inhibition by G6P but not NADP， making the determination of enzyme kinetic parameters complex. We found that the half-saturation concentration of G6PDH isoform 1 is increased under reducing conditions. The other two isoforms exhibit less redox regulation， however， isoform 2 is strongly inhibited by NADPH. Re... More
Glucose-6-phosphate dehydrogenase (G6PDH) can initiate the glucose-6-phosphate (G6P) shunt around the Calvin-Benson cycle. To understand the regulation of flux through this pathway， we have characterized the biochemical parameters and redox regulation of the three functional plastidic isoforms of G6PDH. When purified， recombinant proteins were measured， all three exhibited significant substrate inhibition by G6P but not NADP， making the determination of enzyme kinetic parameters complex. We found that the half-saturation concentration of G6PDH isoform 1 is increased under reducing conditions. The other two isoforms exhibit less redox regulation， however， isoform 2 is strongly inhibited by NADPH. Redox regulation of G6PDH1 can be partially reversed by hydrogen peroxide or protected against by the presence of its substrate， G6P. Overall， our results support the conclusion that G6PDH can have significant activity throughout the day and can be dynamically regulated to allow or prevent flux through the glucose-6-phosphate shunt.
