Determination of glucose-6-phosphatase activity using the glucose dehydrogenase-coupled reaction

Anal Biochem. 1988 Aug 15;173(1):185-9. doi: 10.1016/0003-2697(88)90176-5.

Abstract

We present a method to determine glucose 6-phosphate activity. This assay measures the rate of glucose released in the glucose-6-phosphatase reaction. The glucose is oxidized to beta-D-gluconolactone by glucose dehydrogenase in a coupled reaction that uses NAD(P)+. The determination is rapid, reproducible, and does not require withdrawal, precipitation, centrifugation, or neutralization steps. This method provides a simple resolution to the problem of the nonspecific appearance of Pi, which is especially important in studies of regulation of glucose-6-phosphatase performed in the presence of ATP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Gluconates
  • Glucose 1-Dehydrogenase
  • Glucose Dehydrogenases
  • Glucose-6-Phosphatase / analysis*
  • Lactones
  • Microsomes, Liver / enzymology
  • NADP
  • Oxidation-Reduction
  • Rats

Substances

  • Gluconates
  • Lactones
  • NADP
  • Glucose Dehydrogenases
  • Glucose 1-Dehydrogenase
  • Glucose-6-Phosphatase
  • beta-glucono-1,5-lactone