Determination of glucose-6-phosphatase activity using the glucose dehydrogenase-coupled reaction |
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| Authors: | M Alegre C J Ciudad C Fillat J J Guinovart |
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| Affiliation: | Departament de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, Bellaterra, Spain. |
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| 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. |
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