A continuous spectrophotometric assay for the enzymatic marker glucose 6-phosphatase.
A continuous spectrophotometric assay for glucose 6-phosphatase is described. The method uses glucose dehydrogenase and mutarotase as ancillary enzymes. Glucose 6-phosphatase activity is measured by following NADH formation at 340 nm. The method is linear, at least up to 38 mU in the test which corresponds to a delta E of 0.24 min-1, when the enzyme is assayed in a microsomal fraction. We also discuss the method's suitability for subcellular fractionation. No other continuous assay for this important enzymatic marker of the endoplasmic reticulum is currently available.
Stio
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Dipartimento di Scienze Biochimiche, Università di Firenze, Italy.
Vanni
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Pinzauti
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D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002021:Buffers; D002458:Cell Fractionation; D005952:Glucose-6-Phosphatase / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008862:Microsomes, Liver / Q000201:enzymology; D051381:Rats; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
D015415:Biomarkers; D002021:Buffers; D005952:Glucose-6-Phosphatase