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Cow Brain Glutaminase: Purification, Characterization, And Mechanism Of Action, Fulung John Chiu

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

A simple, quick, and sensitive radiometric assay for glutaminase has been established. This assay uses L-(U-¹⁴C)-glutamine as the substrate and measures the ¹⁴CO₂ produced when the reaction is coupled to glutamate decarboxylase. An efficient purification procedure for cow brain glutaminase has also been developed; the yield is 12%. Steps include acetone extraction, ammonium sulfate fractionation, calcium phosphate gel treatment, and differential gel filtration on Sepharose 4B. The final preparation has a specific activity of 385 μmole/min/mg and shows a single protein band on polyacrylamide gel electrophoresis in sodium dodecyl sulfate; this band corresponds to a …